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| - | <html>
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| - | <div id="kyoto-notebook">
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| - | <div>
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| - | <a name="0715" class="kyoto-jump"></a>
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| - | <h3>7/15</h3>
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| - | <h4>transformation</h4>
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| - | <span class="author">Tatsui, Matsumoto and Sukegawa</span>
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| - | <table class="note">
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| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
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| - | <tr><td>BBa_J23101(Pcon)</td><td>2</td><td>20(2/23)</td><td>22</td><td>LB(CP)</td></tr>
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| - | <tr><td>BBa_B0032(RBS)</td><td>2</td><td>20(2/23)</td><td>22</td><td>LB(CP)</td></tr>
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| - | </table>
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| - | </div>
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| | | | |
| - | <div>
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| - | <a name="0716" class="kyoto-jump"></a>
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| - | <h3>7/16</h3>
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| - | <h4>transformation</h4>
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| - | <span class="author">Tatsui</span>
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| - | <table class="note">
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| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
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| - | <tr><td>BBa_B0030(1)</td><td>2</td><td>20(2/23)</td><td>22</td><td>LB(CP)</td></tr>
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| - | <tr><td>BBa_B0030(2)</td><td>2</td><td>20(bought)</td><td>22</td><td>LB(CP)</td></tr>
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| - | <tr><td>BBa_B0032(1)</td><td>2</td><td>20(2/23)</td><td>22</td><td>LB(CP)</td></tr>
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| - | <tr><td>BBa_B0032(2)</td><td>2</td><td>20(bought)</td><td>22</td><td>LB(CP)</td></tr>
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| - | </table>
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| - | </div>
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| - |
| |
| - | <div>
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| - | <a name="0717" class="kyoto-jump"></a>
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| - | <h3>7/17</h3>
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| - | <h4>Colony PCR</h4>
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| - | <span class="author">Tatsui, Matsumoto, Li, and Yamauchi</span>
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| - | <table class="note">
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| - | <tr><th>NameⅠ</th><th>NameⅡ</th></tr>
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| - | <tr><td>7/16 BBa_B0030(1) Colony_1</td><td>7/16 BBa_B0032(1) Colony_1</td></tr>
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| - | <tr><td>7/16 BBa_B0030(1) Colony_2</td><td>7/16 BBa_B0032(1) Colony_2</td></tr>
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| - | <tr><td>7/16 BBa_B0030(1) Colony_3</td><td>7/16 BBa_B0032(1) Colony_3</td></tr>
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| - | <tr><td>7/16 BBa_B0030(2) Colony_1</td><td>7/16 BBa_B0032(2) Colony_1</td></tr>
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| - | <tr><td>7/16 BBa_B0030(2) Colony_2</td><td>7/16 BBa_B0032(2) Colony_2</td></tr>
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| - | <tr><td>7/16 BBa_B0030(2) Colony_3</td><td>7/16 BBa_B0032(2) Colony_3</td></tr>
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| - | <tr><td>Negative Control</td><td>-</td></tr>
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| - | <tr><td>-</td><td>-</td></tr>
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| - | </table>
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| - |
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| - | <h4>Liquid Culture</h4>
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| - | <span class="author">Tatsui, Matsumoto, Li, and Yamauchi</span>
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| - | <table class="note">
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| - | <tr><th>Name</th><th>medium</th></tr>
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| - | <tr><td>7/16 BBa_B0030(1) Colony_1</td><td>Plusgrow medium(CP)</td></tr>
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| - | <tr><td>7/16 BBa_B0030(1) Colony_2</td><td>Plusgrow medium(CP)</td></tr>
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| - | <tr><td>7/16 BBa_B0030(1) Colony_3</td><td>Plusgrow medium(CP)</td></tr>
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| - | <tr><td>7/16 BBa_B0030(2) Colony_1</td><td>Plusgrow medium(CP)</td></tr>
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| - | <tr><td>7/16 BBa_B0030(2) Colony_2</td><td>Plusgrow medium(CP)</td></tr>
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| - | <tr><td>7/16 BBa_B0030(2) Colony_3</td><td>Plusgrow medium(CP)</td></tr>
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| - | <tr><td>7/16 BBa_B0032(1) Colony_1</td><td>Plusgrow medium(CP)</td></tr>
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| - | <tr><td>7/16 BBa_B0032(1) Colony_2</td><td>Plusgrow medium(CP)</td></tr>
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| - | <tr><td>7/16 BBa_B0032(1) Colony_3</td><td>Plusgrow medium(CP)</td></tr>
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| - | <tr><td>7/16 BBa_B0032(2) Colony_1</td><td>Plusgrow medium(CP)</td></tr>
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| - | <tr><td>7/16 BBa_B0032(2) Colony_2</td><td>Plusgrow medium(CP)</td></tr>
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| - | <tr><td>7/16 BBa_B0032(2) Colony_3</td><td>Plusgrow medium(CP)</td></tr>
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| - | </table>
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| - | </div>
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| - |
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| - | <div>
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| - | <a name="0718" class="kyoto-jump"></a>
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| - | <h3>7/18</h3>
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| - | <h4>Miniprep</h4>
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| - | <span class="author">Tatsui, Matsumoto, Tsujii, Stephane, and Yamauchi</span>
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| - | <table class="note">
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| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
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| - | <tr><td>7/16 BBa_B0030(1) Colony_1</td><td>229</td><td>1.96</td><td>1.40</td></tr>
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| - | <tr><td>7/16 BBa_B0030(1) Colony_2</td><td>209</td><td>1.95</td><td>1.50</td></tr>
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| - | <tr><td>7/16 BBa_B0030(1) Colony_3</td><td>108</td><td>2.12</td><td>2.12</td></tr>
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| - | <tr><td>7/16 BBa_B0030(2) Colony_1</td><td>-</td><td>-</td><td>-</td></tr>
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| - | <tr><td>7/16 BBa_B0030(2) Colony_2</td><td>174</td><td>2.02</td><td>1.56</td></tr>
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| - | <tr><td>7/16 BBa_B0030(2) Colony_3</td><td>158</td><td>2.01</td><td>1.68</td></tr>
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| - | <tr><td>7/16 BBa_B0032(1) Colony_1</td><td>179</td><td>2.00</td><td>2.06</td></tr>
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| - | <tr><td>7/16 BBa_B0032(1) Colony_2</td><td>-</td><td>-</td><td>-</td></tr>
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| - | <tr><td>7/16 BBa_B0032(1) Colony_3</td><td>221</td><td>1.90</td><td>1.37</td></tr>
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| - | <tr><td>7/16 BBa_B0032(2) Colony_1</td><td>225</td><td>1.89</td><td>1.37</td></tr>
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| - | <tr><td>7/16 BBa_B0032(2) Colony_2</td><td>270</td><td>1.98</td><td>1.63</td></tr>
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| - | <tr><td>7/16 BBa_B0032(2) Colony_3</td><td>194</td><td>1.89</td><td>1.33</td></tr>
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| - | </table>
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| - | </div>
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| - |
| |
| - | <div>
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| - | <a name="0730" class="kyoto-jump"></a>
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| - | <h3>7/30</h3>
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| - | <h4>Restriction Enzyme Digestion</h4>
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| - | <span class="author">No name</span>
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| - | <table class="note">
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| - | <tr><th>Sample</th><th>DNA/(µl)</th><th>EcoR1/(µl)</th><th>Xba1/(µl)</th><th>Spe1/(µl)</th><th>Pst1/(µl)</th><th>Buffur/(µl)</th><th>BSA/(µl)</th><th>MilliQ/(µl)</th><th>TOTAL/(µl)</tr>
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| - | <tr><td>7/16 BBa_B0030(2) Colony_1</td><td>1</td><td>0.3</td><td>0</td><td>0</td><td>0</td><td>1(H)</td><td>0</td><td>7.7</td><td>10</td></tr>
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| - | <tr><td>7/16 BBa_B0030(2) Colony_1</td><td>1</td><td>0</td><td>0.3</td><td>0</td><td>0</td><td>1(M)</td><td>1</td><td>6.7</td><td>10</td></tr>
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| - | <tr><td>7/16 BBa_B0030(2) Colony_1</td><td>1</td><td>0</td><td>0</td><td>0.3</td><td>0</td><td>1(M)</td><td>0</td><td>7.7</td><td>10</td></tr>
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| - | <tr><td>7/16 BBa_B0030(2) Colony_1</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0.3</td><td>1(H)</td><td>0</td><td>7.7</td><td>10</td></tr>
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| - | <tr><td>7/16 BBa_B0030(2) Colony_1(Nega)</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1(M)</td><td>0</td><td>8</td><td>10</td></tr>
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| - | </table>
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| - |
| |
| - | <h4>Electrophoresis</h4>
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| - | <span class="author">No name</span>
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| - | <table class="note">
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| - | <tr><th>Lane</th><th>Restriction Enzyme Digestion Products</td></tr>
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| - | <tr><td>1</td><td>7/30 BBa_B0030(2) Colony_1(E CUT)</td></tr>
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| - | <tr><td>2</td><td>7/30 BBa_B0030(2) Colony_1(X CUT)</td></tr>
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| - | <tr><td>3</td><td>7/30 BBa_B0030(2) Colony_1(S CUT)</td></tr>
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| - | <tr><td>4</td><td>7/30 BBa_B0030(2) Colony_1(P CUT)</td></tr>
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| - | <tr><td>5</td><td>7/30 BBa_B0030(2) Colony_1(Nega)</td></tr>
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| - | <tr><td>6</td><td>1kbp ladder</td></tr>
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| - | </table>
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| - | </div>
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| - |
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| - | <div>
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| - | <a name="0731" class="kyoto-jump"></a>
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| - | <h3>7/31</h3>
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| - | <h4>Transformation</h4>
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| - | <span class="author">Hashitani, Tatsui, Li and Yamauchi</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
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| - | <tr><td>BBa_E0240(RBS-GFP-DT)</td><td>2</td><td>20</td><td>22</td><td>LB(CP)</td></tr>
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| - | <tr><td>J23100(Pcon)</td><td>2</td><td>20</td><td>22</td><td>LB(AMP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Liquid Culture</h4>
| |
| - | <span class="author">No name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Name</th><th>medium</th></tr>
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| - | <tr><td>7/31 J23100(Pcon) Colony_1</td><td>Plusgrow medium(AMP)</td></tr>
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| - | <tr><td>7/31 J23100(Pcon) Colony_2</td><td>Plusgrow medium(AMP)</td></tr>
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| - | <tr><td>7/31 J23100(Pcon) Colony_3</td><td>Plusgrow medium(AMP)</td></tr>
| |
| - | <tr><td>7/31 J23100(Pcon) Colony_4</td><td>Plusgrow medium(AMP)</td></tr>
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| - | <tr><td>7/31 J23100(Pcon) Colony_5</td><td>Plusgrow medium(AMP)</td></tr>
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| - | <tr><td>7/31 J23100(Pcon) Colony_6</td><td>Plusgrow medium(AMP)</td></tr>
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| - | </table>
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| - |
| |
| - | <h4>Restriction Enzyme Digestion</h4>
| |
| - | <span class="author">Hashitani, Tatsui, Li and Yamauchi</span>
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| - | <table class="note">
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| - | <tr><th>7/18 RBS Sample</th><th>DNA/(µl)</th><th>EcoR1/(µl)</th><th>Spe1/(µl)</th><th>Pst1/(µl)</th><th>Xba1/(µl)</th><th>Buffur/(µl)</th><th>BSA/(µl)</th><th>MilliQ/(µl)</th><th>TOTAL/(µl)</tr>
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| - | <tr><td>BBa_B0032</td><td>5</td><td>0.3</td><td>0</td><td>0</td><td>0</td><td>1(H)</td><td>0</td><td>3.7</td><td>10</td></tr>
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| - | <tr><td>BBa_B0032</td><td>5</td><td>0</td><td>0.3</td><td>0</td><td>0</td><td>1(M)</td><td>0</td><td>3.7</td><td>10</td></tr>
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| - | <tr><td>BBa_B0032</td><td>5</td><td>0</td><td>0</td><td>0.3</td><td>0</td><td>1(H)</td><td>0</td><td>3.7</td><td>10</td></tr>
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| - | <tr><td>BBa_B0032</td><td>5</td><td>0</td><td>0</td><td>0</td><td>0.3</td><td>1(M)</td><td>1</td><td>2.7</td><td>10</td></tr>
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| - | <tr><td>BBa_B0032(Nega)</td><td>5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1(M)</td><td>0</td><td>4</td><td>10</td></tr>
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| - | </table>
| |
| - | </div>
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| - |
| |
| - | <div>
| |
| - | <a name="0801" class="kyoto-jump"></a>
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| - | <h3>8/1</h3>
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| - | <h4>Electrophoresis</h4>
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| - | <span class="author">Honda, Li and Yamauchi</span>
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| - | <table class="note">
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| - | <tr><th>Lane</th><th>Restriction Enzyme Digestion Products</td></tr>
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| - | <tr><td>1</td><td>1kb ladder</td></tr>
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| - | <tr><td>2</td><td>7/31 BBa_B0032(E CUT)</td></tr>
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| - | <tr><td>3</td><td>7/31 BBa_B0032(S CUT)Error</td></tr>
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| - | <tr><td>4</td><td>-</td></tr>
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| - | <tr><td>5</td><td>-</td></tr>
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| - | <tr><td>6</td><td>7/31 BBa_B0032(P CUT)</td></tr>
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| - | <tr><td>7</td><td>7/31 BBa_B0032(X CUT)</td></tr>
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| - | <tr><td>8</td><td>7/31 BBa_B0032(Nega)</td></tr>
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| - | </table>
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| - |
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| - | <h4>PCR</h4>
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| - | <span class="author">Tatsui</span>
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| - | <table class="note">
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| - | <tr><th>BBa_E0240(RBS-GFP-DT)</th><th>Volume</th></tr>
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| - | <tr><td>10×buffur/(µl)</td><td>5</td></tr>
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| - | <tr><td>2mMdNTPs/(µl)</th><td>5</td></tr>
| |
| - | <tr><td>25mM MgSO4/(µl)</td><td>2</td></tr>
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| - | <tr><td>VF Primer/(µl)</td><td>1.5</td></tr>
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| - | <tr><td>VR Primer/(µl)</td><td>1.5</td></tr>
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| - | <tr><td>template/(µl)</td><td>0.5</td></tr>
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| - | <tr><td>KOD-plus/(µl)</td><td>1</td></tr>
| |
| - | <tr><td>MilliQ/(µl)</td><td>33.5</td></tr>
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| - | <tr><td>TOTAL/(µl)</td><td>50</td></tr>
| |
| - | </table>
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| - |
| |
| - | <table class="note">
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| - | <tr><th>Steps</th><th>Temparature</th><th>Time</th><th>cycle</th></tr>
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| - | <tr><td>Pre Denature</td><td>94℃</td><td>2min</td><td>1time</td></tr>
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| - | <tr><td>Denature</td><td>94℃</td><td>15sec</td><td>30cycle</td></tr>
| |
| - | <tr><td>Annealing</td><td>55℃</td><td>30sec</td><td>30cycle</td></tr>
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| - | <tr><td>Extension</td><td>68℃</td><td>1min</td><td>30cycle</td></tr>
| |
| - |
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
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| - | <tr><td>Product</td><td>80</td><td>1.91</td><td>1.78</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0802" class="kyoto-jump"></a>
| |
| - | <h3>8/2</h3>
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">Tatsui, Honda, Matsumoto, Yamauchi, Li, Tsujii, and Nakamura</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>7/31 J23100(Pcon) Colony_1</td><td>145</td><td>2.11</td><td>3.09</td></tr>
| |
| - | <tr><td>7/31 J23100(Pcon) Colony_2</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>7/31 J23100(Pcon) Colony_3</td><td>160</td><td>2.01</td><td>3.13</td></tr>
| |
| - | <tr><td>7/31 J23100(Pcon) Colony_4</td><td>140</td><td>1.87</td><td>2.70</td></tr>
| |
| - | <tr><td>7/31 J23100(Pcon) Colony_5</td><td>105</td><td>2.39</td><td>5.90</td></tr>
| |
| - | <tr><td>7/31 J23100(Pcon) Colony_6</td><td>180</td><td>1.90</td><td>1.37</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0820" class="kyoto-jump"></a>
| |
| - | <h3>8/20</h3>
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>7/31 BBa_E0240(RBS-GFD-DT)</td><td>2</td><td>20</td><td>22</td><td>LB(CP)</td></tr>
| |
| - | <tr><td>BBa_I732095</td><td>2</td><td>20</td><td>22</td><td>LB(CP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0821" class="kyoto-jump"></a>
| |
| - | <h3>8/21</h3>
| |
| - | <h4>Colony PCR</h4>
| |
| - | <span class="author">Shimazaki</span>
| |
| - | <table class="note">
| |
| - | <tr><th>NameⅠ</th></tr>
| |
| - | <tr><td>8/20 BBa_I732095 Colony_1</td></tr>
| |
| - | <tr><td>8/20 BBa_I732095 Colony_2</td></tr>
| |
| - | <tr><td>8/20 BBa_I732095 Colony_3</td></tr>
| |
| - | <tr><td>Negative control</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Electrophoresis</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>PCR Product</th></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td></tr>
| |
| - | <tr><td>2</td><td>8/21 BBa_I732095 Colony_1</td></tr>
| |
| - | <tr><td>3</td><td>8/21 BBa_I732095 Colony_2</td></tr>
| |
| - | <tr><td>4</td><td>8/21 BBa_I732095 Colony_3</td></tr>
| |
| - | <tr><td>5</td><td>Negative Control</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Liquid Culture</h4>
| |
| - | <span class="author">Honda and Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Name</th><th>medium</th></tr>
| |
| - | <tr><td>8/20 BBa_I732095 Colony_1</td><td>Plusgrow medium(CP)</td></tr>
| |
| - | <tr><td>8/20 BBa_I732095 Colony_2</td><td>Plusgrow medium(CP)</td></tr>
| |
| - | <tr><td>8/20 BBa_I732095 Colony_3</td><td>Plusgrow medium(CP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0821" class="kyoto-jump"></a>
| |
| - | <h3>8/22</h3>
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>8/20 BBa_I732095 Colony_1</td><td>430</td><td>1.68</td><td>2.13</td></tr>
| |
| - | <tr><td>8/20 BBa_I732095 Colony_2</td><td>255</td><td>1.87</td><td>1.81</td></tr>
| |
| - | <tr><td>8/20 BBa_I732095 Colony_3</td><td>215</td><td>1.78</td><td>1.35</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Restriction Enzyme Digestion</h4>
| |
| - | <span class="author">Nakamura</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Sample</th><th>DNA/(µl)</th><th>EcoR1/(µl)</th><th>Spe1/(µl)</th><th>Buffur/(µl)</th><th>MilliQ/(µl)</th><th>TOTAL/(µl)</tr>
| |
| - | <tr><td>8/22 BBa_I732095 Colony_1</td><td>4.7</td><td>1</td><td>1</td><td>3(H)</td><td>20.3</td><td>30</td></tr>
| |
| - | <tr><td>8/22 BBa_I732095 Colony_2</td><td>7.8</td><td>1</td><td>1</td><td>3(H)</td><td>17.2</td><td>30</td></tr>
| |
| - | <tr><td>8/22 BBa_I732095 Colony_3</td><td>9.3</td><td>1</td><td>1</td><td>3(H)</td><td>15.7</td><td>30</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Tatsui and Yamauchi</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>BBa_J04450(pSB3C5)</td><td>2</td><td>20</td><td>22</td><td>LB(CP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0825" class="kyoto-jump"></a>
| |
| - | <h3>8/25</h3>
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Tatsui, Kita and Li</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>BBa_J04450(pSB2K3)</td><td>2</td><td>20</td><td>22</td><td>LB(KAN)</td></tr>
| |
| - | <tr><td>BBa_C0051(pSB1A2)</td><td>2</td><td>20</td><td>22</td><td>LB(AMP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Liquid Culture</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Name</th><th>medium</th></tr>
| |
| - | <tr><td>8/22 BBa_J04450(pSB3C5)</td><td>Plusgrow medium(CP)*5</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0826" class="kyoto-jump"></a>
| |
| - | <h3>8/26</h3>
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">Tsujii, Honda and Matsumoto</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>8/25 BBa_J04450(pSB3C5)</td><td>575</td><td>1.37</td><td>1.37</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Liquid Culture</h4>
| |
| - | <span class="author">Tatsui, Matsumoto and Nakamura</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Name</th><th>medium</th></tr>
| |
| - | <tr><td>8/25 BBa_J04450(pSB2K3)</td><td>Plusgrow medium(KAN)*5</td></tr>
| |
| - | <tr><td>8/25 BBa_J0051(pSB1A2)</td><td>Plusgrow medium(AMP)*2</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0827" class="kyoto-jump"></a>
| |
| - | <h3>8/27</h3>
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">No Name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>8/26 BBa_J04450(pSB2K3)</td><td>1045</td><td>1.43</td><td>1.11</td></tr>
| |
| - | <tr><td>8/26 BBa_J0051(pSB1A2)</td><td>790</td><td>1.39</td><td>0.90</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>BBa_J04450(pSB3T5)</td><td>2</td><td>20</td><td>22</td><td>LB(TET)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0828" class="kyoto-jump"></a>
| |
| - | <h3>8/28</h3>
| |
| - | <h4>Liquid Culture</h4>
| |
| - | <span class="author">Yamauchi</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Name</th><th>medium</th></tr>
| |
| - | <tr><td>8/27 BBa_J04450(pSB3T5)</td><td>Plusgrow medium(TET)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0829" class="kyoto-jump"></a>
| |
| - | <h3>8/29</h3>
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">Matsumoto</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>8/28 BBa_J04450(pSB3T5)</td><td>180</td><td>1.60</td><td>1.67</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>BBa_J04450(pSB4A5)</td><td>2</td><td>20</td><td>22</td><td>LB(AMP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0901" class="kyoto-jump"></a>
| |
| - | <h3>9/1</h3>
| |
| - | <h4>Colony PCR</h4>
| |
| - | <table class="note">
| |
| - | <tr><th>NameⅠ</th></tr>
| |
| - | <tr><td>-</td></tr>
| |
| - | <tr><td>-</td></tr>
| |
| - | <tr><td>-</td></tr>
| |
| - | <tr><td>9/1 pSB4A5 Colony_1</td></tr>
| |
| - | <tr><td>9/1 pSB4A5 Colony_2</td></tr>
| |
| - | <tr><td>Negative Control</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Restriction Enzyme Digestion</h4>
| |
| - | <span class="author">No name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Sample</th><th>DNA/(µl)</th><th>EcoR1/(µl)</th><th>Pst1/(µl)</th><th>Buffur/(µl)</th><th>MilliQ/(µl)</th><th>TOTAL/(µl)</tr>
| |
| - | <tr><td>8/28 Miniprep pSB3T5</td><td>25</td><td>2</td><td>2</td><td>6(H)</td><td>25</td><td>60</td></tr>
| |
| - | <tr><td>Negative control_1</td><td>3.7</td><td>0</td><td>0</td><td>1(H)</td><td>5.3</td><td>10</td></tr>
| |
| - | <tr><td>8/22 Miniprep I732095</td><td>14</td><td>3</td><td>3</td><td>9(H)</td><td>61</td><td>90</td></tr>
| |
| - | <tr><td>Negative control_2</td><td>1.5</td><td>0</td><td>0</td><td>1(H)</td><td>7.5</td><td>10</td></tr>
| |
| - | <tr><td>8/27 Miniprep pSB2K3</td><td>6</td><td>3</td><td>3</td><td>9(H)</td><td>69</td><td>90</td></tr>
| |
| - | <tr><td>Negative control_3</td><td>1</td><td>0</td><td>0</td><td>1(H)</td><td>8</td><td>10</td></tr>
| |
| - | <tr><td>8/2 Miniprep Pcon Colony_3</td><td>20</td><td>1.5</td><td>1.5</td><td>5(H)</td><td>17</td><td>45</td></tr>
| |
| - | <tr><td>Negative control_4</td><td>4</td><td>0</td><td>0</td><td>1(H)</td><td>5</td><td>10</td></tr>
| |
| - | <tr><td>8/2 Miniprep Pcon Colony_1</td><td>20</td><td>1.5</td><td>1.5</td><td>5(H)</td><td>17</td><td>45</td></tr>
| |
| - | <tr><td>Negative control_5</td><td>4</td><td>0</td><td>0</td><td>1(H)</td><td>5</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Liquid Culture</h4>
| |
| - | <span class="author">No name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Name</th><th>medium</th></tr>
| |
| - | <tr><td>8/29 BBa_J04450(pSB4A5) Colony_1</td><td>Plusgrow medium(AMP)*2</td></tr>
| |
| - | <tr><td>8/1 Pcon Colony_1</td><td>Plusgrow medium(AMP)*2</td></tr>
| |
| - | <tr><td>8/27 pSB3T5 Colony_1</td><td>Plusgrow medium(TET)*2</td></tr>
| |
| - | </table>
| |
| - |
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0902" class="kyoto-jump"></a>
| |
| - | <h3>9/2</h3>
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">Honda, Yamauchi and Li</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>8/29 BBa_J04450(pSB4A5) Colony_1</td><td>210</td><td>1.61</td><td>1.18</td></tr>
| |
| - | <tr><td>8/1 Pcon Colony_1</td><td>215</td><td>1.79</td><td>1.87</td></tr>
| |
| - | <tr><td>8/27 pSB3T5 Colony_1</td><td>215</td><td>1.58</td><td>0.92</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Gel Extraction</h4>
| |
| - | <span class="author">Matsumoto</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Restriction Enzyme Digestion ProductⅠ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>9/1 pSB3T5 Colony_1 (Nega)</td><td>10</td></tr>
| |
| - | <tr><td>2</td><td>9/1 pSB3T5 Colony_1 (E+P)(Confirmation)</td><td>5</td></tr>
| |
| - | <tr><td>3</td><td>9/1 pSB3T5 Colony_1 (E+P)(Extraction)</td><td>55</td></tr>
| |
| - | <tr><td>4</td><td>1kb ladder</td><td>3</td></tr>
| |
| - | <tr><td>5</td><td>9/1 I732095 (Nega)</td><td>10</td></tr>
| |
| - | <tr><td>6</td><td>9/1 I732095 (E+P)(Confirmation)</td><td>5</td></tr>
| |
| - | <tr><td>7</td><td>9/1 I732095 (E+P)(Extraction)</td><td>85</td></tr>
| |
| - | <tr><td>8</td><td>1kb ladder</td><td>3</td></tr>
| |
| - | <tr><td>9</td><td>9/1 pSB2K3 (Nega)</td><td>10</td></tr>
| |
| - | <tr><td>10</td><td>9/1 pSB2K3 (E+P)(Confirmation)</td><td>5</td></tr>
| |
| - | <tr><td>11</td><td>9/1 pSB2K3 (E+P)(Extraction)</td><td>85</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Sample</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>9/1 Pcon Colony_3 (E+P)(Confirmation)</td><td>5</td></tr>
| |
| - | <tr><td>2</td><td>9/1 Pcon Colony_3 (E+P)(Extraction)</td><td>20</td></tr>
| |
| - | <tr><td>3</td><td>9/1 Pcon Colony_3 (E+P)(Extraction)</td><td>20</td></tr>
| |
| - | <tr><td>4</td><td>9/1 Pcon Colony_3 (Nega)</td><td>10</td></tr>
| |
| - | <tr><td>5</td><td>1kb ladder</td><td>3</td></tr>
| |
| - | <tr><td>6</td><td>9/1 Pcon Colony_1 (E+P)(Confirmation)</td><td>5</td></tr>
| |
| - | <tr><td>7</td><td>9/1 Pcon Colony_1 (E+P)(Extraction)</td><td>20</td></tr>
| |
| - | <tr><td>8</td><td>9/1 Pcon Colony_1 (E+P)(Extraction)</td><td>20</td></tr>
| |
| - | <tr><td>9</td><td>9/1 Pcon Colony_1 (Nega)</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0903" class="kyoto-jump"></a>
| |
| - | <h3>9/3</h3>
| |
| - | <h4>Concentration Measurement</h4>
| |
| - | <span class="author">Matsumoto, Yamauchi and Li</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>pSB2K3</td><td>40</td><td>1.48</td><td>0.10</td></tr>
| |
| - | <tr><td>Pcon Colony_1</td><td>30</td><td>1.76</td><td>0.62</td></tr>
| |
| - | <tr><td>pSB3T5</td><td>50</td><td>2.35</td><td>0.50</td></tr>
| |
| - | <tr><td>pSB1C3</td><td>65</td><td>2.14</td><td>0.10</td></tr>
| |
| - | <tr><td>Pcon Colony_3</td><td>45</td><td>1.56</td><td>0.58</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Ligation</h4>
| |
| - | <span class="author">No name</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Insert/(µL)</th><th colspan=2>Vector/(µL)</th><th>T4 Ligase/(µl)</th><th>2*Buffer/(µl)</th><th>MilliQ/(µl)</th><th>Total/(µl)</th></tr>
| |
| - | <tr><td>Pcon</td><td>2.1</td><td>pSB1C3</td><td>1.5</td><td>1</td><td>5</td><td>0.4</td><td>10</td></tr>
| |
| - | <tr><td>-(Negative Control)</td><td>0</td><td>pSB1C3</td><td>1.5</td><td>1</td><td>5</td><td>2.5</td><td>10</td></tr>
| |
| - | <tr><td>Pcon</td><td>1.3</td><td>pSB2K3</td><td>2.5</td><td>1</td><td>5</td><td>0.2</td><td>10</td></tr>
| |
| - | <tr><td>-(Negative Control)</td><td>0</td><td>pSB2K3</td><td>2.5</td><td>1</td><td>5</td><td>1.5</td><td>10</td></tr>
| |
| - | <tr><td>Pcon</td><td>10</td><td>pSB3T5</td><td>2.3</td><td>1</td><td>5</td><td>0.7</td><td>10</td></tr>
| |
| - | <tr><td>-(Negative Control)</td><td>0</td><td>pSB3T5</td><td>2.3</td><td>1</td><td>5</td><td>1.7</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Matsumoto</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>9/3 Ligation Pcon+pSB1C3</td><td>3</td><td>15</td><td>18</td><td>LB(CP)</td></tr>
| |
| - | <tr><td>9/3 Ligation pSB1C3(Negative Control)</td><td>8</td><td>10</td><td>18</td><td>LB(CP)</td></tr>
| |
| - | <tr><td>9/3 Ligation Pcon+pSB2K3</td><td>3</td><td>15</td><td>18</td><td>LB(KAN)</td></tr>
| |
| - | <tr><td>9/3 Ligation pSB2K3(Negative Control)</td><td>8</td><td>10</td><td>18</td><td>LB(KAN)</td></tr>
| |
| - | <tr><td>9/3 Ligation Pcon+pSB3T5</td><td>3</td><td>15</td><td>18</td><td>LB(TET)</td></tr>
| |
| - | <tr><td>9/3 Ligation pSB3T5(Negative Control)</td><td>2</td><td>10</td><td>12</td><td>LB(TET)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0904" class="kyoto-jump"></a>
| |
| - | <h3>9/4</h3>
| |
| - | <h4>Colony PCR</h4>
| |
| - | <span class="author">Honda and Yamauchi</span>
| |
| - | <table class="note">
| |
| - | <tr><th>NameⅠ</th><th>NameⅡ</th><th>NameⅢ</th><th>NameⅣ</th></tr>
| |
| - | <tr><td>-</td><td>9/3 pSB1C3(Nega) Colony_7</td><td>9/3 Pcon+pSB3T5 Colony_5</td><td>9/3 Pcon+pSB2K3 Colony_1</td></tr>
| |
| - | <tr><td>-</td><td>9/3 pSB1C3(Nega) Colony_8</td><td>9/3 Pcon+pSB3T5 Colony_6</td><td>9/3 Pcon+pSB2K3 Colony_2</td></tr>
| |
| - | <tr><td>9/3 pSB1C3(Nega) Colony_1</td><td>9/3 Pcon+pSB3T5 Colony_1</td><td>9/3 Pcon+pSB3T5 Colony_7</td><td>9/3 Pcon+pSB2K3 Colony_3</td></tr>
| |
| - | <tr><td>9/3 pSB1C3(Nega) Colony_2</td><td>9/3 Pcon+pSB3T5 Colony_2</td><td>9/3 Pcon+pSB3T5 Colony_8</td><td>9/3 Pcon+pSB2K3 Colony_4</td></tr>
| |
| - | <tr><td>9/3 pSB1C3(Nega) Colony_3</td><td>9/3 Pcon+pSB3T5 Colony_3</td><td>9/3 Pcon+pSB3T5 Colony_9</td><td>9/3 Pcon+pSB2K3 Colony_5</td></tr>
| |
| - | <tr><td>9/3 pSB1C3(Nega) Colony_4</td><td>9/3 Pcon+pSB3T5 Colony_4</td><td>9/3 Pcon+pSB3T5 Colony_10</td><td>9/3 Pcon+pSB2K3 Colony_6</td></tr>
| |
| - | <tr><td>9/3 pSB1C3(Nega) Colony_5</td><td>9/3 pSB1C3(Nega) Colony_9</td><td>9/3 pSB3T5(Nega) Colony_1</td><td>9/3 Pcon+pSB2K3 Colony_7</td></tr>
| |
| - | <tr><td>9/3 pSB1C3(Nega) Colony_6</td><td>9/3 pSB1C3(Nega) Colony_10</td><td>9/3 pSB3T5(Nega) Colony_2</td><td>9/3 Pcon+pSB2K3 Colony_8</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>NameⅤ</th><th>NameⅥ</th><th>NameⅦ</th><th>NameⅧ</th></tr>
| |
| - | <tr><td>9/3 Pcon+pSB2K3 Colony_9</td><td>9/3 Pcon+pSB1C3 Colony_2</td><td>9/3 Pcon+pSB1C3 Colony_10</td><td>9/3 pSB1C3(Nega) Colony_8</td></tr>
| |
| - | <tr><td>9/3 Pcon+pSB2K3 Colony_10</td><td>9/3 Pcon+pSB1C3 Colony_3</td><td>9/3 pSB1C3(Nega) Colony_1</td><td>9/3 pSB1C3(Nega) Colony_9</td></tr>
| |
| - | <tr><td>9/3 pSB2K3(Nega) Colony_1</td><td>9/3 Pcon+pSB1C3 Colony_4</td><td>9/3 pSB1C3(Nega) Colony_2</td><td>9/3 pSB1C3(Nega) Colony_10</td></tr>
| |
| - | <tr><td>9/3 pSB2K3(Nega) Colony_2</td><td>9/3 Pcon+pSB1C3 Colony_5</td><td>9/3 pSB1C3(Nega) Colony_3</td><td>9/3 pSB1C3(Nega) Colony_11</td></tr>
| |
| - | <tr><td>9/3 pSB2K3(Nega) Colony_3</td><td>9/3 Pcon+pSB1C3 Colony_6</td><td>9/3 pSB1C3(Nega) Colony_4</td><td>-</td></tr>
| |
| - | <tr><td>9/3 pSB2K3(Nega) Colony_4</td><td>9/3 Pcon+pSB1C3 Colony_7</td><td>9/3 pSB1C3(Nega) Colony_5</td><td>-</td></tr>
| |
| - | <tr><td>9/3 pSB2K3(Nega) Colony_5</td><td>9/3 Pcon+pSB1C3 Colony_8</td><td>9/3 pSB1C3(Nega) Colony_6</td><td>-</td></tr>
| |
| - | <tr><td>9/3 Pcon+pSB1C3 Colony_1</td><td>9/3 Pcon+pSB1C3 Colony_9</td><td>9/3 pSB1C3(Nega) Colony_7</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>NameⅨ</th><th>NameⅩ</th></tr>
| |
| - | <tr><td>9/3 Pcon+pSB1C3 Colony_11</td><td>8/22 Miniprep I732095 Colony_3</td></tr>
| |
| - | <tr><td>Negative Control</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Restriction Enzyme Digestion</h4>
| |
| - | <span class="author">No name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Sample</th><th>DNA/(µl)</th><th>EcoR1/(µl)</th><th>Pst1/(µl)</th><th>Buffur/(µl)</th><th>MilliQ/(µl)</th><th>TOTAL/(µl)</tr>
| |
| - | <tr><td>8/27 Miniprep pSB2K3</td><td>2</td><td>1</td><td>1</td><td>3(H)</td><td>23</td><td>30</td></tr>
| |
| - | <tr><td>8/27 Miniprep pSB2K3(Nega)</td><td>1</td><td>0</td><td>0</td><td>1(H)</td><td>8</td><td>10</td></tr>
| |
| - | <tr><td>9/2 Miniprep pSB3T5</td><td>10</td><td>1</td><td>1</td><td>3(H)</td><td>15</td><td>30</td></tr>
| |
| - | <tr><td>9/2 Miniprep pSB3T5(Nega)</td><td>3</td><td>0</td><td>0</td><td>1(H)</td><td>6</td><td>10</td></tr>
| |
| - | <tr><td>9/2 Miniprep pSB4A5</td><td>10</td><td>1</td><td>1</td><td>3(H)</td><td>15</td><td>30</td></tr>
| |
| - | <tr><td>9/2 Miniprep pSB4A5(Nega)</td><td>3</td><td>0</td><td>0</td><td>1(H)</td><td>6</td><td>10</td></tr>
| |
| - | <tr><td>8/22 Miniprep pSB1C3 Colony_2</td><td>2</td><td>1</td><td>1</td><td>1(H)</td><td>5</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0905" class="kyoto-jump"></a>
| |
| - | <h3>9/5</h3>
| |
| - | <h4>Electrophoresis</h4>
| |
| - | <span class="author">Honda and Matsumoto</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR ProductsⅠ</td><th>Lane</th><th>Colony PCR ProductsⅠ</td></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td><td>10</td><td>9/4 pSB1C3(Nega) Colony_7</td></tr>
| |
| - | <tr><td>2</td><td>-</td><td>11</td><td>9/4 pSB1C3(Nega) Colony_8</td></tr>
| |
| - | <tr><td>3</td><td>-</td><td>12</td><td>9/4 pSB1C3(Nega) Colony_9</td></tr>
| |
| - | <tr><td>4</td><td>9/4 pSB1C3(Nega) Colony_1</td><td>13</td><td>9/4 pSB1C3(Nega) Colony_10</td></tr>
| |
| - | <tr><td>5</td><td>9/4 pSB1C3(Nega) Colony_2</td><td>14</td><td>9/4 Pcon+pSB2K3 Colony_1</td></tr>
| |
| - | <tr><td>6</td><td>9/4 pSB1C3(Nega) Colony_3</td><td>15</td><td>9/4 Pcon+pSB2K3 Colony_2</td></tr>
| |
| - | <tr><td>7</td><td>9/4 pSB1C3(Nega) Colony_4</td><td>16</td><td>9/4 Pcon+pSB2K3 Colony_3</td></tr>
| |
| - | <tr><td>8</td><td>9/4 pSB1C3(Nega) Colony_5</td><td>17</td><td>1kb ladder</td></tr>
| |
| - | <tr><td>9</td><td>9/4 pSB1C3(Nega) Colony_6</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR ProductsⅡ</td><th>Lane</th><th>Colony PCR ProductsⅡ</td></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td><td>10</td><td>9/4 pSB3T5(Nega) Colony_2</td></tr>
| |
| - | <tr><td>2</td><td>9/4 Pcon+pSB2K3 Colony_4</td><td>11</td><td>9/4 Pcon+pSB2K3 Colony_1</td></tr>
| |
| - | <tr><td>3</td><td>9/4 Pcon+pSB2K3 Colony_5</td><td>12</td><td>9/4 Pcon+pSB2K3 Colony_2</td></tr>
| |
| - | <tr><td>4</td><td>9/4 Pcon+pSB2K3 Colony_6</td><td>13</td><td>9/4 Pcon+pSB2K3 Colony_3</td></tr>
| |
| - | <tr><td>5</td><td>9/4 Pcon+pSB2K3 Colony_7</td><td>14</td><td>9/4 Pcon+pSB2K3 Colony_4</td></tr>
| |
| - | <tr><td>6</td><td>9/4 Pcon+pSB2K3 Colony_8</td><td>15</td><td>9/4 Pcon+pSB2K3 Colony_5</td></tr>
| |
| - | <tr><td>7</td><td>9/4 Pcon+pSB2K3 Colony_9</td><td>16</td><td>9/4 Pcon+pSB2K3 Colony_6</td></tr>
| |
| - | <tr><td>8</td><td>9/4 Pcon+pSB3T5 Colony_10</td><td>17</td><td>1kb ladder</td></tr>
| |
| - | <tr><td>9</td><td>9/4 pSB3T5(Nega) Colony_1</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR ProductsⅢ</td><th>Lane</th><th>Colony PCR ProductsⅢ</td></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td><td>10</td><td>9/4 pSB2K3(Nega) Colony_5</td></tr>
| |
| - | <tr><td>2</td><td>9/4 Pcon+pSB2K3 Colony_7</td><td>11</td><td>9/4 pSB1C3(Nega) Colony_1</td></tr>
| |
| - | <tr><td>3</td><td>9/4 Pcon+pSB2K3 Colony_8</td><td>12</td><td>9/4 pSB1C3(Nega) Colony_2</td></tr>
| |
| - | <tr><td>4</td><td>9/4 Pcon+pSB2K3 Colony_9</td><td>13</td><td>9/4 pSB1C3(Nega) Colony_3</td></tr>
| |
| - | <tr><td>5</td><td>9/4 Pcon+pSB2K3 Colony_10</td><td>14</td><td>9/4 pSB1C3(Nega) Colony_4</td></tr>
| |
| - | <tr><td>6</td><td>9/4 pSB2K3(Nega) Colony_1</td><td>15</td><td>9/4 pSB1C3(Nega) Colony_5</td></tr>
| |
| - | <tr><td>7</td><td>9/4 pSB2K3(Nega) Colony_2</td><td>16</td><td>9/4 pSB1C3(Nega) Colony_6</td></tr>
| |
| - | <tr><td>8</td><td>9/4 pSB2K3(Nega) Colony_3</td><td>17</td><td>1kb ladder</td></tr>
| |
| - | <tr><td>9</td><td>9/4 pSB2K3(Nega) Colony_4</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR ProductsⅣ</td><th>Lane</th><th>Colony PCR ProductsⅣ</td></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td><td>10</td><td>9/4 Pcon+pSB1C3 Colony_5</td></tr>
| |
| - | <tr><td>2</td><td>9/4 pSB1C3(Nega) Colony_7</td><td>11</td><td>9/4 Pcon+pSB1C3 Colony_6</td></tr>
| |
| - | <tr><td>3</td><td>9/4 pSB1C3(Nega) Colony_8</td><td>12</td><td>9/4 Pcon+pSB1C3 Colony_7</td></tr>
| |
| - | <tr><td>4</td><td>9/4 pSB1C3(Nega) Colony_9</td><td>13</td><td>9/4 Pcon+pSB1C3 Colony_8</td></tr>
| |
| - | <tr><td>5</td><td>9/4 pSB1C3(Nega) Colony_10</td><td>14</td><td>9/4 Pcon+pSB1C3 Colony_9</td></tr>
| |
| - | <tr><td>6</td><td>9/4 Pcon+pSB1C3 Colony_1</td><td>15</td><td>9/4 Pcon+pSB1C3 Colony_10</td></tr>
| |
| - | <tr><td>7</td><td>9/4 Pcon+pSB1C3 Colony_2</td><td>16</td><td>8/22 Miniprep BBa_I732095 Colony_3</td></tr>
| |
| - | <tr><td>8</td><td>9/4 Pcon+pSB1C3 Colony_3</td><td>17</td><td>1kb ladder</td></tr>
| |
| - | <tr><td>9</td><td>9/4 Pcon+pSB1C3 Colony_4</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR ProductsⅤ</td></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td></tr>
| |
| - | <tr><td>2</td><td>9/4 Pcon+pSB1C3 Colony_11</td></tr>
| |
| - | <tr><td>3</td><td>9/4 Negative Control</td></tr>
| |
| - | <tr><td>4</td><td>8/22 Miniprep BBa_I732095 Colony_3</td></tr>
| |
| - | <tr><td>5</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>-</td></tr>
| |
| - | <tr><td>7</td><td>-</td></tr>
| |
| - | <tr><td>8</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Gel Extraction</h4>
| |
| - | <span class="author">No Name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Restriction Enzyme Digestion ProductⅠ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td><td>5</td></tr>
| |
| - | <tr><td>2</td><td>9/4 pSB2K3(E+P)</td><td>5</td></tr>
| |
| - | <tr><td>3</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>4</td><td>9/4 pSB2K3(E+P)</td><td>25</td></tr>
| |
| - | <tr><td>5</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>9/4 pSB2K3(Nega)</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Restriction Enzyme Digestion ProductⅡ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td><td>5</td></tr>
| |
| - | <tr><td>2</td><td>9/4 pSB3T5(E+P)</td><td>5</td></tr>
| |
| - | <tr><td>3</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>4</td><td>9/4 pSB3T5(E+P)</td><td>25</td></tr>
| |
| - | <tr><td>5</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>9/4 pSB3T5(Nega)</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Restriction Enzyme Digestion ProductⅢ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td><td>5</td></tr>
| |
| - | <tr><td>2</td><td>9/4 pSB4A5(E+P)</td><td>5</td></tr>
| |
| - | <tr><td>3</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>4</td><td>9/4 pSB4A5(E+P)</td><td>25</td></tr>
| |
| - | <tr><td>5</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>9/4 pSB4A5(Nega)</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Restriction Enzyme Digestion ProductⅣ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td><td>5</td></tr>
| |
| - | <tr><td>2</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>3</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>4</td><td>9/4 pSB1C3(E+P)</td><td>10</td></tr>
| |
| - | <tr><td>5</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/4 pSB2K3(E+P)</td><td>36</td><td>1.25</td><td>0.24</td></tr>
| |
| - | <tr><td>9/4 pSB3T5(E+P)</td><td>10</td><td>1.00</td><td>-/td></tr>
| |
| - | <tr><td>9/4 pSB4A5(E+P)</td><td>40</td><td>-</td><td>0.01</td></tr>
| |
| - | <tr><td>9/4 pSB1C3(E+P)</td><td>75</td><td>1.35</td><td>0.14</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0908" class="kyoto-jump"></a>
| |
| - | <h3>9/8</h3>
| |
| - | <h4>Cloning PCR</h4>
| |
| - | <span class="author">Honda and Li</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Target</th><th>mamB</th><th>mamQ</th><th>mamL</th></tr>
| |
| - | <tr><td>2×KAPA HiFi Hotstart Ready Mix/(µl)</td><td>12.5</td><td>12.5</td><td>12.5</td></tr>
| |
| - | <tr><td>Fw Primer/(µl)</td><td>1.5(mamB)</td><td>1.5(mamQ)</td><td>1.5(mamL)</td></tr>
| |
| - | <tr><td>Rv Primer/(µl)</td><td>1.5(mamB)</td><td>1.5(mamQ)</td><td>1.5(mamL)</td></tr>
| |
| - | <tr><td>AMB-1 Genome/(µl)</td><td>1</td><td>1</td><td>1</td></tr>
| |
| - | <tr><td>MilliQ/(µl)</td><td>10</td><td>10</td><td>10</td></tr>
| |
| - | <tr><td>TOTAL/(µl)</td><td>25</td><td>25</td><td>25</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Steps</th><th>Temparature</th><th>Time</th><th>cycle</th></tr>
| |
| - | <tr><td>Pre Denature</td><td>98℃</td><td>5min</td><td>1time</td></tr>
| |
| - | <tr><td>Denature</td><td>98℃</td><td>20sec</td><td>25cycle</td></tr>
| |
| - | <tr><td>Annealing</td><td>65℃</td><td>15sec</td><td>25cycle</td></tr>
| |
| - | <tr><td>Extension</td><td>72℃</td><td>5min</td><td>25cycle</td></tr>
| |
| - | <tr><td>Final elongation</td><td>72℃</td><td>5min</td><td>1time</td></tr>
| |
| - | <tr><td>Final hold</td><td>4℃</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Ligation</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Insert/(µL)</th><th colspan=2>Vector/(µL)</th><th>T4 Ligase/(µl)</th><th>2*Buffer/(µl)</th><th>MilliQ/(µl)</th><th>Total/(µl)</th></tr>
| |
| - | <tr><td>Pcon</td><td>1</td><td>pSB3T5</td><td>10</td><td>1</td><td>11</td><td>0</td><td>22</td></tr>
| |
| - | <tr><td>-(Negative Control)</td><td>0</td><td>pSB1C3</td><td>3</td><td>0</td><td>3</td><td>0</td><td>6</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Electrophoresis</h4>
| |
| - | <span class="author">Li</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Cloning PCR Products</td></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td></tr>
| |
| - | <tr><td>2</td><td>9/8 Cloning PCR (AMB-1 mamB)</td></tr>
| |
| - | <tr><td>3</td><td>9/8 Cloning PCR (AMB-1 mamQ)</td></tr>
| |
| - | <tr><td>4</td><td>9/8 Cloning PCR (AMB-1 mamL)</td></tr>
| |
| - | <tr><td>5</td><td>1kb ladder</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>9/8 Ligation Pcon+pSB3T5</td><td>3</td><td>30</td><td>33</td><td>LB(TET)</td></tr>
| |
| - | <tr><td>9/8 Ligation pSB3T5(Negative Control)</td><td>3</td><td>30</td><td>33</td><td>LB(TET)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Liquid Culture</h4>
| |
| - | <span class="author">No name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Name</th><th>medium</th></tr>
| |
| - | <tr><td>9/3 Transformation Pcon+pSB1C3 Colony_8</td><td>Plusgrow medium(CP)*2</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0909" class="kyoto-jump"></a>
| |
| - | <h3>9/9</h3>
| |
| - | <h4>DNA Purification</h4>
| |
| - | <span class="author">Li</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/8 Cloning PCR (AMB-1 mamB)</td><td>165</td><td>1.54</td><td>1.57</td></tr>
| |
| - | <tr><td>9/8 Cloning PCR (AMB-1 mamQ)</td><td>250</td><td>1.69</td><td>1.92</td></tr>
| |
| - | <tr><td>9/8 Cloning PCR (AMB-1 mamL)</td><td>190</td><td>1.56</td><td>1.79</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">Li</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/8 Liquid Culture Pcon+pSB1C3</td><td>90</td><td>1.65</td><td>1.17</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Colony PCR</h4>
| |
| - | <span class="author">Honda and Li</span>
| |
| - | <table class="note">
| |
| - | <tr><th>NameⅠ</th></tr>
| |
| - | <tr><td>-</td></tr>
| |
| - | <tr><td>9/8 Transformation Pcon+pSB3T5 Colony_1</td></tr>
| |
| - | <tr><td>9/8 Transformation Pcon+pSB3T5 Colony_2</td></tr>
| |
| - | <tr><td>9/8 Transformation Pcon+pSB3T5 Colony_3</td></tr>
| |
| - | <tr><td>9/8 Transformation Pcon+pSB3T5 Colony_4</td></tr>
| |
| - | <tr><td>9/8 Transformation Pcon+pSB3T5 Colony_5</td></tr>
| |
| - | <tr><td>9/8 Transformation pSB3T5(Nega) Colony_1</td></tr>
| |
| - | <tr><td>Negative control</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Electrophoresis</h4>
| |
| - | <span class="author">Honda and Li</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Cloning PCR Products</td></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td></tr>
| |
| - | <tr><td>2</td><td>9/8 Colony PCR Pcon+pSB3T5 Colony_1</td></tr>
| |
| - | <tr><td>3</td><td>9/8 Colony PCR Pcon+pSB3T5 Colony_2</td></tr>
| |
| - | <tr><td>4</td><td>9/8 Colony PCR Pcon+pSB3T5 Colony_3</td></tr>
| |
| - | <tr><td>5</td><td>9/8 Colony PCR Pcon+pSB3T5 Colony_4</td></tr>
| |
| - | <tr><td>6</td><td>9/8 Colony PCR Pcon+pSB3T5 Colony_5</td></tr>
| |
| - | <tr><td>7</td><td>9/8 Colony PCR pSB3T5(Nega) Colony_1</td></tr>
| |
| - | <tr><td>8</td><td>9/8 Colony PCR Nega</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Restriction Enzyme Digestion</h4>
| |
| - | <span class="author">No name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Sample</th><th>DNA/(µl)</th><th>Pst1/(µl)</th><th>Spe1/(µl)</th><th>Buffur/(µl)</th><th>MilliQ/(µl)</th><th>TOTAL/(µl)</tr>
| |
| - | <tr><td>9/9 Miniprep Pcon+pSB1C3</td><td>22</td><td>1</td><td>1</td><td>3(H)</td><td>3</td><td>30</td></tr>
| |
| - | <tr><td>9/9 Miniprep Pcon+pSB1C3(Nega)</td><td>1</td><td>0</td><td>0</td><td>1(H)</td><td>8</td><td>10</td></tr>
| |
| - | <tr><td>8/2 Miniprep Pcon Colony_4</td><td>14</td><td>1</td><td>1</td><td>3(H)</td><td>11</td><td>30</td></tr>
| |
| - | <tr><td>8/2 Miniprep Pcon Colony_4(Nega)</td><td>1</td><td>0</td><td>0</td><td>1(H)</td><td>8</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0910" class="kyoto-jump"></a>
| |
| - | <h3>9/10</h3>
| |
| - |
| |
| - | <h4>Gel Extraction</h4>
| |
| - | <span class="author">Tatsui and Yamaura</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Restriction Enzyme Digestion ProductⅠ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>2</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>3</td><td>9/10 Digestion Pcon+pSB1C3(S+P)</td><td>-</td></tr>
| |
| - | <tr><td>4</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>5</td><td>9/10 Digestion Pcon+pSB1C3(Nega)</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>7</td><td>Pcon(X+P)</td><td>-</td></tr>
| |
| - | <tr><td>8</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>9</td><td>Pcon(Nega)</td><td>-</td></tr>
| |
| - | <tr><td>10</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>11</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>Pcon(X+P)</td><td>70</td><td>1.40</td><td>0.13</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>PCR</h4>
| |
| - | <span class="author">No Name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Target</th><th>E+X+RBS+mamB+S</th><th>X+RBS+mamL+His+S+P</th><th>X+RBS+mamQ+His+S+P</th></tr>
| |
| - | <tr><td>2×KAPA HiFi Hotstart Ready Mix/(µl)</td><td>12.5</td><td>12.5</td><td>12.5</td></tr>
| |
| - | <tr><td>Fw Primer/(µl)</td><td>0.75(E+X+RBS+mamB)</td><td>0.75(X+RBS+mamL)</td><td>0.75(X+RBS+mamQ)</td></tr>
| |
| - | <tr><td>Rv Primer/(µl)</td><td>0.75(S+mamB)</td><td>0.75(P+S+His+mamL)</td><td>0.75(P+S+His+mamQ)</td></tr>
| |
| - | <tr><td>9/9 DNA Purification Product/(µl)</td><td>Trace</td><td>Trace</td><td>Trace</td></tr>
| |
| - | <tr><td>MilliQ/(µl)</td><td>11</td><td>11</td><td>11</td></tr>
| |
| - | <tr><td>TOTAL/(µl)</td><td>25</td><td>25</td><td>25</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Steps</th><th>Temparature</th><th>Time</th><th>cycle</th></tr>
| |
| - | <tr><td>Pre Denature</td><td>98℃</td><td>5min</td><td>1time</td></tr>
| |
| - | <tr><td>Denature</td><td>98℃</td><td>20sec</td><td>25cycle</td></tr>
| |
| - | <tr><td>Annealing</td><td>60℃</td><td>15sec</td><td>25cycle</td></tr>
| |
| - | <tr><td>Extension</td><td>72℃</td><td>15sec</td><td>25cycle</td></tr>
| |
| - | <tr><td>Final elongation</td><td>72℃</td><td>5min</td><td>1time</td></tr>
| |
| - | <tr><td>Final hold</td><td>12℃</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Electrophoresis</h4>
| |
| - | <span class="author">Li</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Cloning PCR Products</td></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td></tr>
| |
| - | <tr><td>2</td><td>9/10 PCR E+X+RBS+mamB+S(922bp)</td></tr>
| |
| - | <tr><td>3</td><td>9/10 PCR X+RBS+mamL+His+S+P(340bp)</td></tr>
| |
| - | <tr><td>4</td><td>-</td></tr>
| |
| - | <tr><td>5</td><td>1kb ladder</td></tr>
| |
| - | <tr><td>6</td><td>9/10 PCR X+RBS+mamQ+His+S+P(953bp)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>DNA Purification</h4>
| |
| - | <span class="author">Honda and Yamaura</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/10 PCR E+X+RBS+mamB+S</td><td>200</td><td>1.84</td><td>2.15</td></tr>
| |
| - | <tr><td>9/10 PCR X+RBS+mamL+His+S+P</td><td>120</td><td>1.69</td><td>1.84</td></tr>
| |
| - | <tr><td>9/10 PCR X+RBS+mamQ+His+S+P</td><td>200</td><td>1.70</td><td>2.16</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Restriction Enzyme Digestion</h4>
| |
| - | <span class="author">Shimizu</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Sample</th><th>DNA/(µl)</th><th>Xba1/(µl)</th><th>Pst1/(µl)</th><th>Buffur/(µl)</th><th>MilliQ/(µl)</th><th>TOTAL/(µl)</tr>
| |
| - | <tr><td>9/10 PCR E+X+RBS+mamB+S</td><td>10</td><td>1</td><td>1</td><td>3(H)</td><td>15</td><td>30</td></tr>
| |
| - | <tr><td>9/10 PCR X+RBS+mamL+His+S+P</td><td>16.7</td><td>1</td><td>1</td><td>3(H)</td><td>8.3</td><td>30</td></tr>
| |
| - | <tr><td>9/10 PCR X+RBS+mamQ+His+S+P</td><td>10</td><td>1</td><td>1</td><td>3(H)</td><td>15</td><td>30</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Ligation</h4>
| |
| - | <span class="author">Tatsui and Murada</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Insert/(µL)</th><th colspan=2>Vector/(µL)</th><th>T4 Ligase/(µl)</th><th>2*Buffer/(µl)</th><th>MilliQ/(µl)</th><th>Total/(µl)</th></tr>
| |
| - | <tr><td>9/10 DNA Purification mamQ</td><td>1</td><td>YCplac111</td><td>1</td><td>1</td><td>5</td><td>2</td><td>10</td></tr>
| |
| - | <tr><td>9/10 DNA Purification mamL</td><td>1</td><td>YCplac111</td><td>1</td><td>1</td><td>5</td><td>2</td><td>10</td></tr>
| |
| - | <tr><td>9/10 DNA Purification mamB</td><td>1</td><td>YCplac111</td><td>1</td><td>1</td><td>5</td><td>2</td><td>10</td></tr>
| |
| - | <tr><td>Control</td><td>0</td><td>YCplac111</td><td>1</td><td>1</td><td>5</td><td>2</td><td>9</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Tatsui and Murada</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>9/10 Ligation mamQ+YCplac111</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>9/10 Ligation mamL+YCplac111</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>9/10 Ligation mamB+YCplac111</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0911" class="kyoto-jump"></a>
| |
| - | <h3>9/11</h3>
| |
| - | <h4>DNA Purification</h4>
| |
| - | <span class="author">No Name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/10 Digestion mamB(X+P)</td><td>65</td><td>1.68</td><td>0.51</td></tr>
| |
| - | <tr><td>9/10 Digestion mamL(X+P)</td><td>30</td><td>1.95</td><td>0.81</td></tr>
| |
| - | <tr><td>9/10 Digestion mamQ(X+P)</td><td>60</td><td>1.65</td><td>1.22</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Electrophoresis</h4>
| |
| - | <span class="author">Murada</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR Products</td></tr>
| |
| - | <tr><td></td><td></td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR Products</td></tr>
| |
| - | <tr><td></td><td></td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR Products</td></tr>
| |
| - | <tr><td></td><td></td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR Products</td></tr>
| |
| - | <tr><td></td><td></td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR Products</td></tr>
| |
| - | <tr><td></td><td></td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Ligation</h4>
| |
| - | <span class="author">No Name</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Insert/(µL)</th><th colspan=2>Vector/(µL)</th><th>T4 Ligase/(µl)</th><th>2*Buffer/(µl)</th><th>MilliQ/(µl)</th><th>Total/(µl)</th></tr>
| |
| - | <tr><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Yamauchi</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>9/11 Ligation mamL</td><td>2</td><td>10</td><td>12</td><td>-</td></tr>
| |
| - | <tr><td>9/11 Ligation mamQ</td><td>1</td><td>5</td><td>6</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Liquid Culture</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Name</th><th>medium</th></tr>
| |
| - | <tr><td>9/8 Master Pcon+pSB1C3 Colony_3</td><td>Plusgrow medium(CP)</td></tr>
| |
| - | <tr><td>9/10 Transformation mamL+YCplac111</td><td>Plusgrow medium(AMP)</td></tr>
| |
| - | <tr><td>9/10 Transformation mamQ+YCplac111</td><td>Plusgrow medium(AMP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0912" class="kyoto-jump"></a>
| |
| - | <h3>9/12</h3>
| |
| - | <h4>Colony PCR</h4>
| |
| - | <span class="author">Yamaura</span>
| |
| - | <table class="note">
| |
| - | <tr><th>NameⅠ</th></tr>
| |
| - | <tr><td></td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Electrophoresis</h4>
| |
| - | <span class="author">Yamaura</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR Products</td></tr>
| |
| - | <tr><td></td><td></td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/11 Liquid Culture Pcon+pSB1C3</td><td>110</td><td>1.83</td><td>2.63</td></tr>
| |
| - | <tr><td>9/11 Liquid Culture mamL+YCplac111</td><td>300</td><td>1.75</td><td>2.38</td></tr>
| |
| - | <tr><td>9/11 Liquid Culture mamQ+YCplac111</td><td>160</td><td>1.81</td><td>2.37</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Restriction Enzyme Digestion</h4>
| |
| - | <span class="author">Yasuda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Sample</th><th>DNA/(µl)</th><th>Xba1/(µl)</th><th>Pst1/(µl)</th><th>Spe1/(µl)</th><th>Buffur/(µl)</th><th>BSA</th><th>MilliQ/(µl)</th><th>TOTAL/(µl)</tr>
| |
| - | <tr><td>9/12 Miniprep mamL+YCplac111</td><td>6.7</td><td>1</td><td>1</td><td>0</td><td>3(H)</td><td>3</td><td>15.3</td><td>30</td></tr>
| |
| - | <tr><td>9/12 Miniprep mamQ+YCplac111</td><td>12.5</td><td>1</td><td>1</td><td>0</td><td>3(H)</td><td>3</td><td>9.5</td><td>30</td></tr>
| |
| - | <tr><td>9/12 Miniprep Pcon+pSB1C3</td><td>18.1</td><td>0</td><td>1</td><td>1</td><td>3(H)</td><td>0</td><td>6.9</td><td>30</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Ligation</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Insert/(µL)</th><th colspan=2>Vector/(µL)</th><th>T4 Ligase/(µl)</th><th>2*Buffer/(µl)</th><th>MilliQ/(µl)</th><th>Total/(µl)</th></tr>
| |
| - | <tr><td>9/12 Digestion mamB(E+S)</td><td>3</td><td>9/8 Digestion pSB1C3(E+S)</td><td>1</td><td>1</td><td>5</td><td>0</td><td>10</td></tr>
| |
| - | <tr><td>9/10 mamB</td><td>0.5</td><td>YCplac111</td><td>1</td><td>1</td><td>5</td><td>2.5</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>DNA Purification</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>mamB(E+S)</td><td>50</td><td>1.75</td><td>2.04</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>9/12 Ligation mamB+pSB1C3</td><td>1</td><td>10</td><td>11</td><td>LB(CP)</td></tr>
| |
| - | <tr><td>9/12 Ligation mamB+YCplac111</td><td>1</td><td>10</td><td>11</td><td>LB(AMP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Colony PCR</h4>
| |
| - | <span class="author">Li</span>
| |
| - | <table class="note">
| |
| - | <tr><th>NameⅠ</th><th>NameⅡ</th></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Colony_1</td><td>9/11 Transformation mamL+pSB1A2 Colony_9</td></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Colony_2</td><td>9/11 Transformation mamL+pSB1A2 Colony_10</td></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Colony_3</td><td>9/11 Transformation mamQ+pSB1A2 Colony_1</td></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Colony_4</td><td>9/11 Transformation mamQ+pSB1A2 Colony_2</td></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Colony_5</td><td>9/11 Transformation mamQ+pSB1A2 Colony_3</td></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Colony_6</td><td>9/11 Transformation mamQ+pSB1A2 Colony_4</td></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Colony_7</td><td>9/11 Transformation mamQ+pSB1A2 Colony_5</td></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Colony_8</td><td>9/11 Transformation mamQ+pSB1A2 Colony_6</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Electrophoresis</h4>
| |
| - | <span class="author">Li</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR ProductsⅠ</td><th>Lane</th><th>Colony PCR ProductsⅠ</td></tr>
| |
| - | <tr><td>1</td><td>9/12 mamL+pSB1A2 Colony_1</td><td>10</td><td>9/12 mamL+pSB1A2 Colony_9</td></tr>
| |
| - | <tr><td>2</td><td>9/12 mamL+pSB1A2 Colony_2</td><td>11</td><td>9/12 mamL+pSB1A2 Colony_10</td></tr>
| |
| - | <tr><td>3</td><td>9/12 mamL+pSB1A2 Colony_3</td><td>12</td><td>9/12 mamQ+pSB1A2 Colony_1</td></tr>
| |
| - | <tr><td>4</td><td>9/12 mamL+pSB1A2 Colony_4</td><td>13</td><td>9/12 mamQ+pSB1A2 Colony_2</td></tr>
| |
| - | <tr><td>5</td><td>9/12 mamL+pSB1A2 Colony_5</td><td>14</td><td>9/12 mamQ+pSB1A2 Colony_3</td></tr>
| |
| - | <tr><td>6</td><td>9/12 mamL+pSB1A2 Colony_6</td><td>15</td><td>9/12 mamQ+pSB1A2 Colony_4</td></tr>
| |
| - | <tr><td>7</td><td>9/12 mamL+pSB1A2 Colony_7</td><td>16</td><td>9/12 mamQ+pSB1A2 Colony_5</td></tr>
| |
| - | <tr><td>8</td><td>9/12 mamL+pSB1A2 Colony_8</td><td>17</td><td>9/12 mamQ+pSB1A2 Colony_6</td></tr>
| |
| - | <tr><td>9</td><td>1kb ladder</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Colony PCR</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>NameⅠ</th></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Green_1</td></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Green_2</td></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Green_3</td></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Green_4</td></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Green_5</td></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Black_7</td></tr>
| |
| - | <tr><td>9/11 Transformation mamL+pSB1A2 Black_8</td></tr>
| |
| - | <tr><td>9/11 Transformation mamQ+pSB1A2 Black_1</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Electrophoresis</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR ProductsⅠ</td></tr>
| |
| - | <tr><td>1</td><td>9/12 mamL+pSB1A2 Green_1</td></tr>
| |
| - | <tr><td>2</td><td>9/12 mamL+pSB1A2 Green_2</td></tr>
| |
| - | <tr><td>3</td><td>9/12 mamL+pSB1A2 Green_3</td></tr>
| |
| - | <tr><td>4</td><td>9/12 mamL+pSB1A2 Green_4</td></tr>
| |
| - | <tr><td>5</td><td>9/12 mamL+pSB1A2 Green_5</td></tr>
| |
| - | <tr><td>6</td><td>9/12 mamL+pSB1A2 Black_7</td></tr>
| |
| - | <tr><td>7</td><td>9/12 mamL+pSB1A2 Black_8</td></tr>
| |
| - | <tr><td>8</td><td>9/12 mamQ+pSB1A2 Black_1</td></tr>
| |
| - | <tr><td>9</td><td>1kb ladder</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Liquid Culture</h4>
| |
| - | <span class="author">Yamauchi</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Name</th><th>medium</th></tr>
| |
| - | <tr><td>9/11 Transformation mamQ+pSB1A2 Colony_1</td><td>Plusgrow medium(AMP)</td></tr>
| |
| - | <tr><td>9/11 Transformation mamQ+pSB1A2 Colony_2</td><td>Plusgrow medium(AMP)</td></tr>
| |
| - | <tr><td>9/11 Transformation mamQ+pSB1A2 Colony_3</td><td>Plusgrow medium(AMP)</td></tr>
| |
| - | <tr><td>9/11 Transformation mamQ+pSB1A2 Colony_4</td><td>Plusgrow medium(AMP)</td></tr>
| |
| - | <tr><td>9/11 Transformation mamQ+pSB1A2 Colony_5</td><td>Plusgrow medium(AMP)</td></tr>
| |
| - | <tr><td>9/11 Transformation mamQ+pSB1A2 Colony_6</td><td>Plusgrow medium(AMP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0913" class="kyoto-jump"></a>
| |
| - | <h3>9/13</h3>
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/12 Liquid Culture mamQ+pSB1A2 Colony_1 Error</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>9/12 Liquid Culture mamQ+pSB1A2 Colony_2 Error</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>9/12 Liquid Culture mamQ+pSB1A2 Colony_3 Error</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>9/12 Liquid Culture mamQ+pSB1A2 Colony_4</td><td>80</td><td>2.02</td><td>2.76</td></tr>
| |
| - | <tr><td>9/12 Liquid Culture mamQ+pSB1A2 Colony_5</td><td>70</td><td>2.13</td><td>3.41</td></tr>
| |
| - | <tr><td>9/12 Liquid Culture mamQ+pSB1A2 Colony_6</td><td>110</td><td>1.96</td><td>1.70</td></tr>
| |
| - | <tr><td>9/12 Liquid Culture Pcon+pSB1C3</td><td>110</td><td>1.96</td><td>1.70</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>PCR Labeling and Sequence</h4>
| |
| - | <span class="author">No Name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Target</th><th>9/13 mamQ+pSB1A2 Colony_4</th><th>9/13 mamQ+pSB1A2 Colony_5</th><th>9/13 mamQ+pSB1A2 Colony_6</th><th>Pcon+pSB1A2</th><th>9/13 mamQ+pSB1A2 Colony_4</th><th>9/13 mamQ+pSB1A2 Colony_5</th><th>9/13 mamQ+pSB1A2 Colony_6</th></tr>
| |
| - | <tr><td>Template/(µl)</td><td>5</td><td>4</td><td>6</td><td>4</td><td>5</td><td>6</td><td>4</td></tr>
| |
| - | <tr><td>Big dye/(µl)</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td></tr>
| |
| - | <tr><td>5×Buffer/(µl)</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td></tr>
| |
| - | <tr><td>Primer/(µl)</td><td>0.5(VF)</td><td>0.5(VF)</td><td>0.5(VF)</td><td>0.5(VF)</td><td>0.5(VR)</td><td>0.5(VR)</td><td>0.5(VR)</td></tr>
| |
| - | <tr><td>MilliQ/(µl)</td><td>2.75</td><td>1.75</td><td>3.75</td><td>3.75</td><td>2.75</td><td>1.75</td><td>3.75</td></tr>
| |
| - | <tr><td>TOTAL/(µl)</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Target</th><th>9/12 mamQ+YCplac111</th><th>9/12 mamL+YCplac111</th><th>9/12 mamQ+YCplac111</th><th>9/12 mamL+YCplac111</th><th>9/12 mamQ+YCplac111</th></tr>
| |
| - | <tr><td>Template/(µl)</td><td>3</td><td>1.5</td><td>3</td><td>1.5</td><td>3</td></tr>
| |
| - | <tr><td>Big dye/(µl)</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td></tr>
| |
| - | <tr><td>5×Buffer/(µl)</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td></tr>
| |
| - | <tr><td>Primer/(µl)</td><td>0.5(M13F)</td><td>0.5(M13F)</td><td>0.5(M13R)</td><td>0.5(M13R)</td><td>0.5(seqpLQB-2)</td></tr>
| |
| - | <tr><td>MilliQ/(µl)</td><td>4.75</td><td>6.25</td><td>4.75</td><td>6.25</td><td>4.75</td></tr>
| |
| - | <tr><td>TOTAL/(µl)</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Gel Extraction</h4>
| |
| - | <span class="author">No Name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Restriction Enzyme Digestion ProductⅠ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>2</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>3</td><td>9/12 Digestion mamQ+YCpla111(X+P)(992bp)</td><td>-</td></tr>
| |
| - | <tr><td>4</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>5</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>7</td><td>9/12 Digestion mamL+YCpla111(X+P)(340bp)</td><td>-</td></tr>
| |
| - | <tr><td>8</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>9</td><td>9/12 Digestion Pcon+pSB1C3(Nega)(5600bp)</td><td>-</td></tr>
| |
| - | <tr><td>10</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>11</td><td>9/12 Digestion Pcon+pSB1C3(S+P)(2070bp)</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/12 Digestion mamL+YCpla111(X+P)</td><td>10</td><td>1.00</td><td>0.70</td></tr>
| |
| - | <tr><td>9/12 Digestion Pcon+pSB1C3(S+P)</td><td>150</td><td>1.25</td><td>0.71</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Restriction Enzyme Digestion</h4>
| |
| - | <span class="author">Li</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Sample</th><th>DNA/(µl)</th><th>Xba1/(µl)</th><th>Spe1/(µl)</th><th>Pst1/(µl)</th><th>EcoR1/(µl)</th><th>Buffur/(µl)</th><th>BSA</th><th>MilliQ/(µl)</th><th>TOTAL/(µl)</tr>
| |
| - | <tr><td>9/13 Miniprep mamQ+pSB1A2 Colony_4</td><td>12.5</td><td>0.5</td><td>0</td><td>0.5</td><td>0</td><td>1.7(M)</td><td>1.6</td><td>0</td><td>16.8</td></tr>
| |
| - | <tr><td>9/13 Miniprep mamQ+pSB1A2 Colony_5</td><td>14.3</td><td>0.5</td><td>0</td><td>0.5</td><td>0</td><td>1.9(M)</td><td>1.86</td><td>0</td><td>19.06</td></tr>
| |
| - | <tr><td>9/13 Miniprep mamQ+pSB1A2 Colony_6</td><td>9.1</td><td>0.5</td><td>0</td><td>0.5</td><td>0</td><td>1.5(M)</td><td>1.5</td><td>1.9</td><td>15</td></tr>
| |
| - | <tr><td>9/13 Miniprep mamQ+pSB1A2 Colony_6(Nega)</td><td>3</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0.5(M)</td><td>0.5</td><td>1</td><td>5</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Ligation</h4>
| |
| - | <span class="author">Yasuda</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Insert/(µL)</th><th colspan=2>Vector/(µL)</th><th>T4 Ligase/(µl)</th><th>2*Buffer/(µl)</th><th>MilliQ/(µl)</th><th>Total/(µl)</th></tr>
| |
| - | <tr><td>9/13 Gel Extraction mamL(X+P)</td><td>3</td><td>Pcon+pSB1C3(S+P)</td><td>1</td><td>1</td><td>5</td><td>0</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/12 Liquid Culture Pcon+pSB1C3</td><td>240</td><td>1.78</td><td>1.59</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0914" class="kyoto-jump"></a>
| |
| - | <h3>9/14</h3>
| |
| - | <h4>Gel Extraction</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Restriction Enzyme Digestion ProductⅠ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td><td>3</td></tr>
| |
| - | <tr><td>2</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>3</td><td>9/13 mamQ Colony_4(X+P)</td><td>16.8</td></tr>
| |
| - | <tr><td>4</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>5</td><td>1kb ladder</td><td>3</td></tr>
| |
| - | <tr><td>6</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>7</td><td>9/13 mamQ Colony_5(X+P)</td><td>19.06</td></tr>
| |
| - | <tr><td>8</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>9</td><td>9/13 mamQ Colony_6(Nega)</td><td>5</td></tr>
| |
| - | <tr><td>10</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>11</td><td>9/13 mamQ Colony_6(X+P)</td><td>15</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Ligation</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Insert/(µL)</th><th colspan=2>Vector/(µL)</th><th>T4 Ligase/(µl)</th><th>2*Buffer/(µl)</th><th>MilliQ/(µl)</th><th>Total/(µl)</th></tr>
| |
| - | <tr><td>mamB(E+S)</td><td>4</td><td>pSB1C3(E+S)</td><td>3</td><td>1</td><td>8</td><td>0</td><td>16</td></tr>
| |
| - | <tr><td>mamL(X+P)</td><td>3.3</td><td>pSB1C3(E+S)</td><td>0.7</td><td>1</td><td>5</td><td>0</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>PCR</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Target</th><th>E+X+RBS+mamB+S</th></tr>
| |
| - | <tr><td>2×KAPA HiFi Hotstart Ready Mix/(µl)</td><td>12.5</td></tr>
| |
| - | <tr><td>Fw Primer/(µl)</td><td>0.75(E+X+RBS+mamB)</td>/tr>
| |
| - | <tr><td>Rv Primer/(µl)</td><td>0.75(S+mamB)</td></tr>
| |
| - | <tr><td>9/9 DNA Purification Product/(µl)</td><td>Trace</td></tr>
| |
| - | <tr><td>MilliQ/(µl)</td><td>11</td></tr>
| |
| - | <tr><td>TOTAL/(µl)</td><td>25</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Steps</th><th>Temparature</th><th>Time</th><th>cycle</th></tr>
| |
| - | <tr><td>Pre Denature</td><td>98℃</td><td>5min</td><td>1time</td></tr>
| |
| - | <tr><td>Denature</td><td>98℃</td><td>20sec</td><td>25cycle</td></tr>
| |
| - | <tr><td>Annealing</td><td>60℃</td><td>15sec</td><td>25cycle</td></tr>
| |
| - | <tr><td>Extension</td><td>72℃</td><td>15sec</td><td>25cycle</td></tr>
| |
| - | <tr><td>Final elongation</td><td>72℃</td><td>5min</td><td>1time</td></tr>
| |
| - | <tr><td>Final hold</td><td>12℃</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>9/14 Ligation Pcon+mamL+pSB1C3</td><td>1</td><td>10</td><td>11</td><td>LB(CP)</td></tr>
| |
| - | <tr><td>9/14 Ligation Pcon+mamL+pSB1C3(Nega)</td><td>1</td><td>10</td><td>11</td><td>LB(CP)</td></tr>
| |
| - | <tr><td>9/14 Ligation mamB+pSB1C3</td><td>1</td><td>10</td><td>11</td><td>LB(CP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0915" class="kyoto-jump"></a>
| |
| - | <h3>9/15</h3>
| |
| - | <h4>Colony PCR</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>NameⅠ</th><th>NameⅡ</th></tr>
| |
| - | <tr><td>9/14 Transformation mamB+pSB1C3 Colony_1</td><td>9/14 Transformation mamB+pSB1C3 Colony_9</td></tr>
| |
| - | <tr><td>9/14 Transformation mamB+pSB1C3 Colony_2</td><td>9/14 Transformation mamB+pSB1C3 Colony_10</td></tr>
| |
| - | <tr><td>9/14 Transformation mamB+pSB1C3 Colony_3</td><td>9/14 Transformation mamB+pSB1C3 Colony_11</td></tr>
| |
| - | <tr><td>9/14 Transformation mamB+pSB1C3 Colony_4</td><td>9/14 Transformation mamB+pSB1C3 Colony_12</td></tr>
| |
| - | <tr><td>9/14 Transformation mamB+pSB1C3 Colony_5</td><td>9/14 Transformation mamB+pSB1C3 Colony_13</td></tr>
| |
| - | <tr><td>9/14 Transformation mamB+pSB1C3 Colony_6</td><td>9/14 Transformation mamB+pSB1C3 Colony_14</td></tr>
| |
| - | <tr><td>9/14 Transformation mamB+pSB1C3 Colony_7</td><td>9/14 Transformation mamB+pSB1C3 Colony_15</td></tr>
| |
| - | <tr><td>9/14 Transformation mamB+pSB1C3 Colony_8</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Electrophoresis</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR ProductsⅠ</td></tr>
| |
| - | <tr><td>1</td><td>9/14 mamB+pSB1C3 Colony_1</td></tr>
| |
| - | <tr><td>2</td><td>9/14 mamB+pSB1C3 Colony_2</td></tr>
| |
| - | <tr><td>3</td><td>9/14 mamB+pSB1C3 Colony_3</td></tr>
| |
| - | <tr><td>4</td><td>9/14 mamB+pSB1C3 Colony_4</td></tr>
| |
| - | <tr><td>5</td><td>9/14 mamB+pSB1C3 Colony_5</td></tr>
| |
| - | <tr><td>6</td><td>9/14 mamB+pSB1C3 Colony_6</td></tr>
| |
| - | <tr><td>7</td><td>9/14 mamB+pSB1C3 Colony_7</td></tr>
| |
| - | <tr><td>8</td><td>9/14 mamB+pSB1C3 Colony_8</td></tr>
| |
| - | <tr><td>9</td><td>1kb ladder</td></tr>
| |
| - | <tr><td>10</td><td>9/14 mamB+pSB1C3 Colony_9</td></tr>
| |
| - | <tr><td>11</td><td>9/14 mamB+pSB1C3 Colony_10</td></tr>
| |
| - | <tr><td>12</td><td>9/14 mamB+pSB1C3 Colony_11</td></tr>
| |
| - | <tr><td>13</td><td>9/14 mamB+pSB1C3 Colony_12</td></tr>
| |
| - | <tr><td>14</td><td>9/14 mamB+pSB1C3 Colony_13</td></tr>
| |
| - | <tr><td>15</td><td>9/14 mamB+pSB1C3 Colony_14</td></tr>
| |
| - | <tr><td>16</td><td>9/14 mamB+pSB1C3 Colony_15</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Colony PCR</h4>
| |
| - | <span class="author">No Name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>NameⅠ</th></tr>
| |
| - | <tr><td>9/14 Transformation Pcon+mamL+SB1C3 Colony_1</td></tr>
| |
| - | <tr><td>9/14 Transformation mamB+pSB1C3 Colony_9</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>mamB+YCplac111</td><td>1</td><td>10</td><td>11</td><td>LB(AMP)</td></tr>
| |
| - | <tr><td>Pcon+mamL+pSB1C3</td><td>1</td><td>10</td><td>11</td><td>LB(CP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>mamB+pSB1C3 Colony_9</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>mamB+pSB1C3 Colony_10</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>mamB+pSB1C3 Colony_11</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>mamL+pSB1A2 Colony_1</td><td>170</td><td>1.74</td><td>1.41</td></tr>
| |
| - | <tr><td>mamL+pSB1A2 Colony_2</td><td>140</td><td>1.95</td><td>1.43</td></tr>
| |
| - | <tr><td>mamL+pSB1A2 Colony_3</td><td>135</td><td>1.95</td><td>1.60</td></tr>
| |
| - | <tr><td>mamL+pSB1A2 Colony_4</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>mamL+pSB1A2 Colony_5</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>mamL+pSB1A2 Colony_6</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Liquid Culture</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Name</th><th>medium</th></tr>
| |
| - | <tr><td>9/14 Transformation Pcon+mamL+pSB1C3 Colony_1</td><td>Plusgrow medium(CP)</td></tr>
| |
| - | <tr><td>9/14 Transformation mamB+pSB1C3 Colony_9</td><td>Plusgrow medium(CP)</td></tr>
| |
| - | <tr><td>9/14 Transformation mamB+pSB1C3 Colony_10</td><td>Plusgrow medium(CP)</td></tr>
| |
| - | <tr><td>9/14 Transformation mamB+pSB1C3 Colony_11</td><td>Plusgrow medium(CP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Restriction Enzyme Digestion</h4>
| |
| - | <span class="author">Li</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Sample</th><th>DNA/(µl)</th><th>EcoR1/(µl)</th><th>Xba1/(µl)</th><th>Spe1/(µl)</th><th>Pst1/(µl)</th><th>BSA</th><th>MilliQ/(µl)</th><th>Buffur/(µl)</th><th>TOTAL/(µl)</tr>
| |
| - | <tr><td>9/15 DNA Purification mamB</td><td>25</td><td>1</td><td>0</td><td>1</td><td>0</td><td>0</td><td>0</td><td>3(H)</td><td>30</td></tr>
| |
| - | <tr><td>9/15 Mniprep mamL+pSB1A2 Colony_1</td><td>11.8</td><td>0</td><td>1</td><td>0</td><td>1</td><td>3</td><td>10.2</td><td>3(M)</td><td>30</td></tr>
| |
| - | <tr><td>9/15 Mniprep mamL+pSB1A2 Colony_1(Nega)</td><td>0.6</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td><td>8.4</td><td>1(M)</td><td>10</td></tr>
| |
| - | <tr><td>9/15 Mniprep mamL+pSB1A2 Colony_2</td><td>14.3</td><td>0</td><td>1</td><td>0</td><td>1</td><td>3</td><td>7.7</td><td>3(M)</td><td>30</td></tr>
| |
| - | <tr><td>9/15 Mniprep mamL+pSB1A2 Colony_2(Nega)</td><td>0.7</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td><td>8.3</td><td>1(M)</td><td>10</td></tr>
| |
| - | <tr><td>9/15 Mniprep mamL+pSB1A2 Colony_3</td><td>14.8</td><td>0</td><td>1</td><td>0</td><td>1</td><td>3</td><td>7.2</td><td>3(M)</td><td>30</td></tr>
| |
| - | <tr><td>9/15 Mniprep mamL+pSB1A2 Colony_3(Nega)</td><td>0.7</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td><td>8.3</td><td>1(M)</td><td>10</td></tr>
| |
| - | <tr><td>9/15 Mniprep mamL+pSB1A2 Colony_4</td><td>13.3</td><td>0</td><td>1</td><td>0</td><td>1</td><td>3</td><td>8.7</td><td>3(M)</td><td>30</td></tr>
| |
| - | <tr><td>9/15 Mniprep mamL+pSB1A2 Colony_4(Nega)</td><td>0.7</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td><td>8.3</td><td>1(M)</td><td>10</td></tr>
| |
| - | <tr><td>9/13 Pcon+pSB1C3</td><td>20</td><td>0</td><td>0</td><td>1</td><td>1</td><td>0</td><td>5</td><td>3(H)</td><td>30</td></tr>
| |
| - | <tr><td>9/13 Pcon+pSB1C3(Nega)</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>6</td><td>3(H)</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>PCR</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Target</th><th>mamB Colony_10</th><th>mamB Colony_11</th></tr>
| |
| - | <tr><td>2×PrimeSTAR Max Premix/(µl)</td><td>25</td><td>25</td></tr>
| |
| - | <tr><td>Fw mamB Mutation Primer/(µl)</td><td>1</td><td>1</td></tr>
| |
| - | <tr><td>Rv mamB Mutation Primer/(µl)</td><td>1</td><td>1</td></tr>
| |
| - | <tr><td>9/15 Miniprep mamB+pSB1C3/(µl)</td><td>Colony_10(Trace)</td><td>Colony_11(Trace)</td></tr>
| |
| - | <tr><td>MilliQ/(µl)</td><td>23</td><td>23</td></tr>
| |
| - | <tr><td>TOTAL/(µl)</td><td>50</td><td>50</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Steps</th><th>Temparature</th><th>Time</th><th>cycle</th></tr>
| |
| - | <tr><td>Pre Denature</td><td>98℃</td><td>2min</td><td>1time</td></tr>
| |
| - | <tr><td>Denature</td><td>98℃</td><td>10sec</td><td>30cycle</td></tr>
| |
| - | <tr><td>Annealing</td><td>55℃</td><td>15sec</td><td>30cycle</td></tr>
| |
| - | <tr><td>Extension</td><td>72℃</td><td>15sec</td><td>30cycle</td></tr>
| |
| - | <tr><td>Final elongation</td><td>72℃</td><td>5min</td><td>1time</td></tr>
| |
| - | <tr><td>Final hold</td><td>12℃</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0916" class="kyoto-jump"></a>
| |
| - | <h3>9/16</h3>
| |
| - | <h4>PCR Labeling and Sequence</h4>
| |
| - | <span class="author">No Name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Target</th><th>9/15 Pcon+mamL+pSB1C3</th><th>9/15 mamL+pSB1A2 Colony_1</th><th>9/15 mamL+pSB1A2 Colony_2</th><th>9/15 mamL+pSB1A2 Colony_3</th><th>9/15 mamL+pSB1A2 Colony_4</th><th>9/15 mamB+pSB1C3 Colony_9</th><th>9/15 mamB+pSB1C3 Colony_10</th><th>9/15 mamB+pSB1C3 Colony_11</th></tr>
| |
| - | <tr><td>Template/(µl)</td><td>1.8</td><td>2.35</td><td>2.9</td><td>3.0</td><td>3.0</td><td>1.14</td><td>1.2</td><td>1.2</td></tr>
| |
| - | <tr><td>Big dye/(µl)</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td></tr>
| |
| - | <tr><td>5×Buffer/(µl)</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td></tr>
| |
| - | <tr><td>Primer(VF)/(µl)</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td></tr>
| |
| - | <tr><td>MilliQ/(µl)</td><td>5.95</td><td>5.4</td><td>4.85</td><td>4.75</td><td>4.75</td><td>6.61</td><td>6.55</td><td>6.55</td></tr>
| |
| - | <tr><td>TOTAL/(µl)</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Target</th><th>9/15 Pcon+mamL+pSB1C3</th><th>9/15 mamL+pSB1A2 Colony_1</th><th>9/15 mamL+pSB1A2 Colony_2</th><th>9/15 mamL+pSB1A2 Colony_3</th><th>9/15 mamB+pSB1C3 Colony_9</th><th>9/15 mamB+pSB1C3 Colony_10</th><th>9/15 mamB+pSB1C3 Colony_11</th></tr>
| |
| - | <tr><td>Template/(µl)</td><td>1.8</td><td>2.35</td><td>2.9</td><td>3.0</td><td>1.14</td><td>1.2</td><td>1.2</td></tr>
| |
| - | <tr><td>Big dye/(µl)</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td></tr>
| |
| - | <tr><td>5×Buffer/(µl)</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td></tr>
| |
| - | <tr><td>Primer(VR)/(µl)</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td></tr>
| |
| - | <tr><td>MilliQ/(µl)</td><td>5.95</td><td>5.4</td><td>4.85</td><td>4.75</td><td>6.61</td><td>6.55</td><td>6.55</td></tr>
| |
| - | <tr><td>TOTAL/(µl)</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Steps</th><th>Temparature</th><th>Time</th><th>cycle</th></tr>
| |
| - | <tr><td>Pre Denature</td><td>96℃</td><td>2min</td><td>1time</td></tr>
| |
| - | <tr><td>Denature</td><td>96℃</td><td>10sec</td><td>30cycle</td></tr>
| |
| - | <tr><td>Annealing</td><td>55℃</td><td>5sec</td><td>30cycle</td></tr>
| |
| - | <tr><td>Extension</td><td>72℃</td><td>4min</td><td>30cycle</td></tr>
| |
| - | <tr><td>Final elongation</td><td>72℃</td><td>1min</td><td>1time</td></tr>
| |
| - | <tr><td>Final hold</td><td>4℃</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Gel Extraction</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Restriction Enzyme Digestion ProductⅠ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>2</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>3</td><td>9/15 mamL Colony_1(X+P)</td><td>-</td></tr>
| |
| - | <tr><td>4</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>5</td><td>9/15 mamL Colony_2(Nega)</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>7</td><td>9/15 mamL Colony_2(X+P)</td><td>-</td></tr>
| |
| - | <tr><td>8</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>9</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>10</td><td>9/15 mamL Colony_3(Nega)</td><td>-</td></tr>
| |
| - | <tr><td>11</td><td>9/15 mamL Colony_3(X+P)</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Restriction Enzyme Digestion ProductⅡ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>2</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>3</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>4</td><td>9/15 mamQ(X+P)</td><td>-</td></tr>
| |
| - | <tr><td>5</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>9/15 mamQ(Nega)</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>mamL Colony_1</td><td>45</td><td>1.69</td><td>0.60</td></tr>
| |
| - | <tr><td>mamL Colony_2</td><td>60</td><td>1.33</td><td>0.70</td></tr>
| |
| - | <tr><td>mamL Colony_3</td><td>40</td><td>1.76</td><td>0.08</td></tr>
| |
| - | <tr><td>mamQ</td><td>65</td><td>1.35</td><td>0.83</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Dephosphorylation</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th></tr>
| |
| - | <tr><td>9/15 Digestion Pcon+pSB1C3(S+P)</td><td>-</td></tr>
| |
| - | <tr><td>9/15 Gel Extraction Pcon+pSB1C3(S+P)</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>DNA Purification</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/15 Mutation PCR mamB+pSB1C3 Colony_10</td><td>225</td><td>1.64</td><td>1.74</td></tr>
| |
| - | <tr><td>9/15 Mutation PCR mamB+pSB1C3 Colony_11</td><td>205</td><td>1.59</td><td>1.76</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Gel Extraction</h4>
| |
| - | <span class="author">Yamauchi</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>SampleⅠ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>2</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>3</td><td>9/15 Digestion Pcon+pSB1C3(S+P)</td><td>-</td></tr>
| |
| - | <tr><td>4</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>5</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>7</td><td>9/15 Gel Extraction Pcon+pSB1C3(S+P)</td><td>-</td></tr>
| |
| - | <tr><td>8</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>9</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>10</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>11</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Honda and Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>9/16 DNA Purification mamB+pSB1C3 Colony_10</td><td>1</td><td>10</td><td>11</td><td>LB(CP)</td></tr>
| |
| - | <tr><td>9/16 DNA Purification mamB+pSB1C3 Colony_11</td><td>1</td><td>10</td><td>11</td><td>LB(CP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Restriction Enzyme Digestion</h4>
| |
| - | <span class="author">No Name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Sample</th><th>DNA/(µl)</th><th>EcoR1/(µl)</th><th>Xba1/(µl)</th><th>Spe1/(µl)</th><th>Pst1/(µl)</th><th>BSA</th><th>MilliQ/(µl)</th><th>Buffur/(µl)</th><th>TOTAL/(µl)</tr>
| |
| - | <tr><td>9/15 mamQ+pSB1A2 Colony_4</td><td>26.7</td><td>0</td><td>2</td><td>0</td><td>2</td><td>6</td><td>17.3</td><td>6(M)</td><td>60</td></tr>
| |
| - | <tr><td>9/15 mamQ+pSB1A2 Colony_4(Nega)</td><td>0.7</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td><td>7.3</td><td>1(M)</td><td>10</td></tr>
| |
| - | <tr><td>9/15 Pcon+mamL+pSB1C3</td><td>15</td><td>0</td><td>0</td><td>2</td><td>2</td><td>0</td><td>35</td><td>6(H)</td><td>60</td></tr>
| |
| - | <tr><td>9/15 Pcon+mamL+pSB1C3(Nega)</td><td>0.5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>8.5</td><td>1(H)</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>PCR</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Target</th><th>E+Pcon+X+RBS+mamB+His+S(*2)</th></tr>
| |
| - | <tr><td>2×KAPA HiFi Hotstart Ready Mix/(µl)</td><td>12.5</td></tr>
| |
| - | <tr><td>Fw E+Pcon+X+RBS+mamB/(µl)</td><td>0.75</td></tr>
| |
| - | <tr><td>Rv S+His+mamB/(µl)</td><td>0.75</td></tr>
| |
| - | <tr><td>9/9 DNA Purification mamB/(µl)</td><td>Trace</td></tr>
| |
| - | <tr><td>MilliQ/(µl)</td><td>11</td></tr>
| |
| - | <tr><td>TOTAL/(µl)</td><td>25</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/15 mamB+pSB1C3 Colony_9</td><td>350</td><td>1.68</td><td>1.81</td></tr>
| |
| - | <tr><td>9/15 mamB+pSB1C3 Colony_10</td><td>325</td><td>1.65</td><td>1.50</td></tr>
| |
| - | <tr><td>9/15 mamB+pSB1C3 Colony_11</td><td>340</td><td>1.67</td><td>1.68</td></tr>
| |
| - | <tr><td>Pcon+mamL+pSB1C3</td><td>220</td><td>1.75</td><td>2.09</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0917" class="kyoto-jump"></a>
| |
| - | <h3>9/17</h3>
| |
| - | <h4>Colony PCR</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>NameⅠ</th></tr>
| |
| - | <tr><td>9/16 Transformation Mutation mamB Colony_11_1</td></tr>
| |
| - | <tr><td>9/16 Transformation Mutation mamB Colony_11_2</td></tr>
| |
| - | <tr><td>9/16 Transformation Mutation mamB Colony_11_3</td></tr>
| |
| - | <tr><td>9/16 Transformation Mutation mamB Colony_11_4</td></tr>
| |
| - | <tr><td>9/16 Transformation Mutation mamB Colony_10_1</td></tr>
| |
| - | <tr><td>9/16 Transformation Mutation mamB Colony_10_2</td></tr>
| |
| - | <tr><td>9/16 Transformation Mutation mamB Colony_10_3</td></tr>
| |
| - | <tr><td>9/16 Transformation Mutation mamB Colony_10_4</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">Tatsui and Yamauchi</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/15 Pcon+pSB1C3 Colony_3</td><td>110</td><td>1.66</td><td>1.48</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Gel Extraction</h4>
| |
| - | <span class="author">Yamauchi</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>SampleⅠ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>9/16 Mutation PCR Pcon+mamB+His</td><td>-</td></tr>
| |
| - | <tr><td>2</td><td>Pcon+mamL(Nega)</td><td>-</td></tr>
| |
| - | <tr><td>3</td><td>Pcon+mamL(S+P)</td><td>-</td></tr>
| |
| - | <tr><td>4</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>5</td><td>9/16 Mutation PCR Pcon+mamB+His</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>7</td><td>mamQ+pSB1A2 Colony_4(X+P)</td><td>-</td></tr>
| |
| - | <tr><td>8</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>9</td><td>mamQ(Nega)</td><td>-</td></tr>
| |
| - | <tr><td>10</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>11</td><td>mamQ+pSB1A2 Colony_4(X+P)</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>Pcon+mamB+His</td><td>245</td><td>1.87</td><td>2.10</td></tr>
| |
| - | <tr><td>mamQ+pSB1A2 Colony_4(X+P)</td><td>35</td><td>1.87</td><td>0.580</td></tr>
| |
| - | <tr><td>Pcon+mamL(S+P)</td><td>285 Error</td><td>1.14</td><td>0.85</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Ligation</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Insert/(µL)</th><th colspan=2>Vector/(µL)</th><th>T4 Ligase/(µl)</th><th>2*Buffer/(µl)</th><th>MilliQ/(µl)</th><th>Total/(µl)</th></tr>
| |
| - | <tr><td>mamQ(X+P)</td><td>3</td><td>Pcon+mamL(S+P)</td><td>1</td><td>1</td><td>5</td><td>0</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>9/17 Ligation Pcon+mamL+mamQ+pSB1C3</td><td>1</td><td>10</td><td>11</td><td>LB(CP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Restriction Enzyme Digestion</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Sample</th><th>DNA/(µl)</th><th>EcoR1/(µl)</th><th>Spe1/(µl)</th><th>Xba1/(µl)</th><th>Pst1/(µl)</th><th>BSA</th><th>Buffur/(µl)</th><th>MilliQ/(µl)</th><th>TOTAL/(µl)</tr>
| |
| - | <tr><td>9/17 Gel Extraction Pcon+mamB+His</td><td>16.3</td><td>2</td><td>2</td><td>0</td><td>0</td><td>0</td><td>6(H)</td><td>33.7</td><td>60</td></tr>
| |
| - | <tr><td>9/17 Gel Extraction Pcon+mamB+His(Nega)</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1(H)</td><td>8</td><td>10</td></tr>
| |
| - | <tr><td>9/17 Miniprep Pcon+pSB1C3</td><td>36.4</td><td>0</td><td>2</td><td>0</td><td>2</td><td>0</td><td>6(H)</td><td>13.6</td><td>60</td></tr>
| |
| - | <tr><td>9/17 Miniprep Pcon+pSB1C3(Nega)</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1(H)</td><td>8</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Liquid Culture</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Name</th><th>medium</th></tr>
| |
| - | <tr><td>Mutation mamB Colony_10_3</td><td>Plusgrow medium(CP)</td></tr>
| |
| - | <tr><td>Mutation mamB Colony_10_4 </td><td>Plusgrow medium(CP)</td></tr>
| |
| - | <tr><td>Mutation mamB Colony_11_3</td><td>Plusgrow medium(CP)</td></tr>
| |
| - | <tr><td>Mutation mamB Colony_11_4</td><td>Plusgrow medium(CP)</td></tr>
| |
| - | <tr><td>mamQ+pSB1A2 Colony_4</td><td>Plusgrow medium(AMP)*2</td></tr>
| |
| - | <tr><td>Pcon+mamL Colony_1</td><td>Plusgrow medium(CP)*3.5</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0918" class="kyoto-jump"></a>
| |
| - | <h3>9/18</h3>
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">Tatsui and Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/17 Liquid Culture Mutation mamB Colony_10_3</td><td>355</td><td>1.69</td><td>2.02</td></tr>
| |
| - | <tr><td>9/17 Liquid Culture Mutation mamB Colony_10_4</td><td>265</td><td>1.52</td><td>1.55</td></tr>
| |
| - | <tr><td>9/17 Liquid Culture Mutation mamB Colony_11_3</td><td>355</td><td>1.62</td><td>1.77</td></tr>
| |
| - | <tr><td>9/17 Liquid Culture Mutation mamB Colony_11_4</td><td>250</td><td>1.62</td><td>1.51</td></tr>
| |
| - | <tr><td>9/17 Liquid Culture mamQ+pSB1A2 Colony_4</td><td>290</td><td>1.41</td><td>1.10</td></tr>
| |
| - | <tr><td>9/17 Liquid Culture Pcon+mamL+pSB1C3 Colony_1</td><td>85</td><td>1.47</td><td>1.56</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>PCR Labeling and Sequence</h4>
| |
| - | <span class="author">No Name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Target</th><th>9/18 Miniprep Mutation mamB Colony_10_3</th><th>9/18 Miniprep Mutation mamB Colony_10_4</th><th>9/18 Miniprep Mutation mamB Colony_11_3</th><th>9/18 Miniprep Mutation mamB Colony_11_4</th><th>9/18 Miniprep Mutation mamB Colony_10_3</th><th>9/18 Miniprep Mutation mamB Colony_10_4</th><th>9/18 Miniprep Mutation mamB Colony_11_3</th><th>9/18 Miniprep Mutation mamB Colony_11_4</th></tr>
| |
| - | <tr><td>Template/(µl)</td><td>1.1</td><td>1.5</td><td>1.1</td><td>1.6</td><td>1.1</td><td>1.5</td><td>1.1</td><td>1.6</td></tr>
| |
| - | <tr><td>Big dye/(µl)</td><td>2</td><td>2</td><td>2</td><td>2</td><td>2</td><td>2</td><td>2</td><td>2</td></tr>
| |
| - | <tr><td>5×Buffer/(µl)</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
| |
| - | <tr><td>Primer/(µl)</td><td>0.16(VF)</td><td>0.16(VF)</td><td>0.16(VF)</td><td>0.16(VF)</td><td>0.16(VR)</td><td>0.16(VR)</td><td>0.16(VR)</td><td>0.16(VR)</td></tr>
| |
| - | <tr><td>MilliQ/(µl)</td><td>5.74</td><td>5.34</td><td>5.74</td><td>5.24</td><td>5.74</td><td>5.34</td><td>5.74</td><td>5.24</td></tr>
| |
| - | <tr><td>TOTAL/(µl)</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Target</th><th>9/18 Miniprep Mutation mamB Colony_10_3</th><th>9/18 Miniprep Mutation mamB Colony_10_4</th><th>9/18 Miniprep Mutation mamB Colony_11_3</th><th>9/18 Miniprep Mutation mamB Colony_11_4</th></tr>
| |
| - | <tr><td>Template/(µl)</td><td>1.1</td><td>1.5</td><td>1.1</td><td>1.6</td></tr>
| |
| - | <tr><td>Big dye/(µl)</td><td>2</td><td>2</td><td>2</td><td>2</td></tr>
| |
| - | <tr><td>5×Buffer/(µl)</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
| |
| - | <tr><td>Primer/(µl)</td><td>0.16(pQLB seq_2)</td><td>0.16(pQLB seq_2)</td><td>0.16(pQLB seq_2)</td><td>0.16(pQLB seq_2)</td></tr>
| |
| - | <tr><td>MilliQ/(µl)</td><td>5.74</td><td>5.34</td><td>5.74</td><td>5.24</td></tr>
| |
| - | <tr><td>TOTAL/(µl)</td><td>10</td><td>10</td><td>10</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Steps</th><th>Temparature</th><th>Time</th><th>cycle</th></tr>
| |
| - | <tr><td>Pre Denature</td><td>96℃</td><td>2min</td><td>1time</td></tr>
| |
| - | <tr><td>Denature</td><td>96℃</td><td>10sec</td><td>30cycle</td></tr>
| |
| - | <tr><td>Annealing</td><td>50℃</td><td>5sec</td><td>30cycle</td></tr>
| |
| - | <tr><td>Extension</td><td>60℃</td><td>4min</td><td>30cycle</td></tr>
| |
| - | <tr><td>Final elongation</td><td>4℃</td><td>1min</td><td>1time</td></tr>
| |
| - | <tr><td>Final hold</td><td>4℃</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Colony PCR</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>NameⅠ</th><th>NameⅡ</th></tr>
| |
| - | <tr><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_1</td><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_9</td></tr>
| |
| - | <tr><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_2</td><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_10</td></tr>
| |
| - | <tr><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_3</td><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_11</td></tr>
| |
| - | <tr><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_4</td><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_12</td></tr>
| |
| - | <tr><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_5</td><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_13</td></tr>
| |
| - | <tr><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_6</td><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_14</td></tr>
| |
| - | <tr><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_7</td><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_15</td></tr>
| |
| - | <tr><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_8</td><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3 Colony_16</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Electrophoresis</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR ProductsⅠ</td></tr>
| |
| - | <tr><td>1</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_1</td></tr>
| |
| - | <tr><td>2</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_2</td></tr>
| |
| - | <tr><td>3</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_3</td></tr>
| |
| - | <tr><td>4</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_4</td></tr>
| |
| - | <tr><td>5</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_5</td></tr>
| |
| - | <tr><td>6</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_6</td></tr>
| |
| - | <tr><td>7</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_7</td></tr>
| |
| - | <tr><td>8</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_8</td></tr>
| |
| - | <tr><td>9</td><td>1kb ladder</td></tr>
| |
| - | <tr><td>10</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_9</td></tr>
| |
| - | <tr><td>11</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_10</td></tr>
| |
| - | <tr><td>12</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_11</td></tr>
| |
| - | <tr><td>13</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_12</td></tr>
| |
| - | <tr><td>14</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_13</td></tr>
| |
| - | <tr><td>15</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_14</td></tr>
| |
| - | <tr><td>16</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_15</td></tr>
| |
| - | <tr><td>17</td><td>9/18 Pcon+mamL+mamQ+pSB1C3 Colony_16</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>DNA Purification</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/17 Digestion Pcon+mamB+His(E+S)</td><td>165</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Gel Extraction</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>SampleⅠ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>2</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>3</td><td>9/17 Digestion Pcon+pSB1C3(Nega)</td><td>-</td></tr>
| |
| - | <tr><td>4</td><td>9/17 Digestion Pcon+pSB1C3(S+P)</td><td>-</td></tr>
| |
| - | <tr><td>5</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>SampleⅡ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>2</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>3</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>4</td><td>9/17 Digestion Pcon+pSB1C3(S+P)</td><td>-</td></tr>
| |
| - | <tr><td>5</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/17 Digestion Pcon+pSB1C3</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Ligation</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Insert/(µL)</th><th colspan=2>Vector/(µL)</th><th>T4 Ligase/(µl)</th><th>10*Buffer/(µl)</th><th>MilliQ/(µl)</th><th>Total/(µl)</th></tr>
| |
| - | <tr><td>mamQ(X+P)</td><td>6</td><td>9/18 Gel Extraction Pcon+pSB1C3(S+P)</td><td>Trace</td><td>0.5</td><td>1</td><td>2.5</td><td>10</td></tr>
| |
| - | <tr><td>9/18 DNA Purification Pcon+mamB+His(E+S)</td><td>5</td><td>9/14 pSB3T5(E+S)</td><td>2</td><td>0.5</td><td>1</td><td>1.5</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Transformation</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th colspan=2>Sample/(µL)</th><th>CompetentCells/(µl)</th><th>Total/(µl)</th><th>Medium</th></tr>
| |
| - | <tr><td>9/18 Ligation Pcon+mamQ+pSB1C3</td><td>1</td><td>10</td><td>11</td><td>LB(CP)</td></tr>
| |
| - | <tr><td>9/18 Ligation Pcon+mamB+His+pSB3T5</td><td>1</td><td>10</td><td>11</td><td>LB(TET)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Liquid Culture</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Name</th><th>medium</th></tr>
| |
| - | <tr><td>pSB3T5</td><td>Plusgrow medium(TET)*2</td></tr>
| |
| - | <tr><td>9/17 Transformation Pcon+mamL+mamQ+pSB1C3</td><td>Plusgrow medium(CP)</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Restriction Enzyme Digestion</h4>
| |
| - | <span class="author">No Name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Sample</th><th>DNA/(µl)</th><th>EcoR1/(µl)</th><th>Xba1/(µl)</th><th>Spe1/(µl)</th><th>Pst1/(µl)</th><th>BSA</th><th>Buffur/(µl)</th><th>MilliQ/(µl)</th><th>TOTAL/(µl)</tr>
| |
| - | <tr><td>9/18 Miniprep Mutation mamB Colony_10_3</td><td>11.2</td><td>0</td><td>2</td><td>0</td><td>2</td><td>0.8</td><td>8(D)</td><td>56</td><td>80</td></tr>
| |
| - | <tr><td>9/18 Miniprep Mutation mamB Colony_10_3(Nega)</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1(D)</td><td>8</td><td>10</td></tr>
| |
| - | <tr><td>9/18 Miniprep Mutation mamB Colony_10_4</td><td>15.1</td><td>0</td><td>2</td><td>0</td><td>2</td><td>0.8</td><td>8(D)</td><td>52.1</td><td>80</td></tr>
| |
| - | <tr><td>9/18 Miniprep Mutation mamB Colony_10_4(Nega)</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1(D)</td><td>8</td><td>10</td></tr>
| |
| - | <tr><td>9/18 Miniprep Mutation mamB Colony_11_3</td><td>11.2</td><td>0</td><td>2</td><td>0</td><td>2</td><td>0.8</td><td>8(D)</td><td>56</td><td>80</td></tr>
| |
| - | <tr><td>9/18 Miniprep Mutation mamB Colony_11_3(Nega)</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1(D)</td><td>8</td><td>10</td></tr>
| |
| - | <tr><td>9/18 Miniprep Mutation mamB Colony_11_4</td><td>16</td><td>0</td><td>2</td><td>0</td><td>2</td><td>0.8</td><td>8(D)</td><td>51.2</td><td>80</td></tr>
| |
| - | <tr><td>9/18 Miniprep Mutation mamB Colony_11_4(Nega)</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1(D)</td><td>8</td><td>10</td></tr>
| |
| - | <tr><td>9/18 Miniprep mamQ+pSB1A2</td><td>34.5</td><td>0</td><td>5</td><td>0</td><td>5</td><td>2</td><td>20(D)</td><td>133.5</td><td>200</td></tr>
| |
| - | <tr><td>9/18 Miniprep mamQ+pSB1A2(Nega)</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1(D)</td><td>8</td><td>10</td></tr>
| |
| - | <tr><td>9/17 Miniprep Pcon+pSB1C3</td><td>40</td><td>0</td><td>0</td><td>2</td><td>2</td><td>0.8</td><td>8(B)</td><td>27.2</td><td>80</td></tr>
| |
| - | <tr><td>9/17 Miniprep Pcon+pSB1C3(Nega)</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1(B)</td><td>8</td><td>10</td></tr>
| |
| - | <tr><td>9/18 Miniprep Pcon+mamL+pSB1C3</td><td>47</td><td>0</td><td>0</td><td>2</td><td>2</td><td>0.8</td><td>8(B)</td><td>20.2</td><td>80</td></tr>
| |
| - | <tr><td>9/18 Miniprep Pcon+mamL+pSB1C3(Nega)</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1(B)</td><td>8</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | </div>
| |
| - |
| |
| - | <div>
| |
| - | <a name="0919" class="kyoto-jump"></a>
| |
| - | <h3>9/19</h3>
| |
| - | <h4>Miniprep</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/18 Liquid Culture pSB3T5</td><td>115</td><td>1.52</td><td>1.22</td></tr>
| |
| - | <tr><td>9/18 Liquid Culture Pcon+mamL+mamQ+pSB1C3</td><td>75</td><td>1.59</td><td>1.20</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>PCR Labeling and Sequence</h4>
| |
| - | <span class="author">No Name</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Target</th><th>9/19 Miniprep Pcon+mamL+mamQ+pSB1C3</th><th>9/19 Miniprep Pcon+mamL+mamQ+pSB1C3</th><th>9/19 Miniprep Pcon+mamL+mamQ+pSB1C3</th><th>9/19 Miniprep Pcon+mamL+mamQ+pSB1C3</th></tr>
| |
| - | <tr><td>Template/(µl)</td><td>3.5</td><td>3.5</td><td>3.5</td><td>3.5</td></tr>
| |
| - | <tr><td>Big dye/(µl)</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td></tr>
| |
| - | <tr><td>5×Buffer/(µl)</td><td>1.75</td><td>1.75</td><td>1.75</td><td>1.75</td></tr>
| |
| - | <tr><td>Primer/(µl)</td><td>0.5(VF)</td><td>0.5(VR)</td><td>0.5(seq_1)</td><td>0.5(seq_2)</td></tr>
| |
| - | <tr><td>MilliQ/(µl)</td><td>4.25</td><td>4.25</td><td>4.25</td><td>4.25</td></tr>
| |
| - | <tr><td>TOTAL/(µl)</td><td>10.5</td><td>10.5</td><td>10.5</td><td>10.5</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Steps</th><th>Temparature</th><th>Time</th><th>cycle</th></tr>
| |
| - | <tr><td>Pre Denature</td><td>96℃</td><td>2min</td><td>1time</td></tr>
| |
| - | <tr><td>Denature</td><td>96℃</td><td>10sec</td><td>30cycle</td></tr>
| |
| - | <tr><td>Annealing</td><td>50℃</td><td>5sec</td><td>30cycle</td></tr>
| |
| - | <tr><td>Extension</td><td>60℃</td><td>4min</td><td>30cycle</td></tr>
| |
| - | <tr><td>Final elongation</td><td>4℃</td><td>1min</td><td>1time</td></tr>
| |
| - | <tr><td>Final hold</td><td>4℃</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Gel Extraction</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>SampleⅠ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>2</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>3</td><td>9/18 Digestion Mutation mamB Colony_10_3(X+P)</td><td>40</td></tr>
| |
| - | <tr><td>4</td><td>9/18 Digestion Mutation mamB Colony_10_3(Nega)</td><td>10</td></tr>
| |
| - | <tr><td>5</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>7</td><td>9/18 Digestion Mutation mamB Colony_10_3(X+P)</td><td>40</td></tr>
| |
| - | <tr><td>8</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>9</td><td>1kb ladder</td><td>-</td></tr>
| |
| - | <tr><td>10</td><td>9/18 Digestion Mutation mamB Colony_10_4(Nega)</td><td>10</td></tr>
| |
| - | <tr><td>11</td><td>9/18 Digestion Mutation mamB Colony_10_4(X+P)</td><td>40</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>SampleⅡ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>9/18 Digestion mamQ+pSB1A2(X+P)</td><td>200</td></tr>
| |
| - | <tr><td>2</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>3</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>4</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>5</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>6</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>SampleⅢ</th><th>Volume/(µl)</th></tr>
| |
| - | <tr><td>1</td><td>100b ladder</td><td>-</td></tr>
| |
| - | <tr><td>2</td><td>9/18 Digestion Pcon+pSB1C3(Nega)</td><td>10</td></tr>
| |
| - | <tr><td>3</td><td>9/18 Digestion Pcon+pSB1C3(S+P)</td><td>80</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>mamB Colony_10_3(X+P)</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>mamB Colony_10_4(X+P)</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>mamQ+pSB1A2(X+P)</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | <tr><td>Pcon+pSB1C3(S+P)</td><td>-</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Colony PCR</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>NameⅠ</th></tr>
| |
| - | <tr><td>9/18 Transformation Pcon+mamQ+pSB1C3 Colony_1</td></tr>
| |
| - | <tr><td>9/18 Transformation Pcon+mamQ+pSB1C3 Colony_2</td></tr>
| |
| - | <tr><td>9/18 Transformation Pcon+mamQ+pSB1C3 Colony_3</td></tr>
| |
| - | <tr><td>9/18 Transformation Pcon+mamQ+pSB1C3 Colony_4</td></tr>
| |
| - | <tr><td>9/18 Transformation Pcon+mamQ+pSB1C3 Colony_5</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Electrophoresis</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Lane</th><th>Colony PCR ProductsⅠ</td></tr>
| |
| - | <tr><td>1</td><td>-</td></tr>
| |
| - | <tr><td>2</td><td>-</td></tr>
| |
| - | <tr><td>3</td><td>Pcon+mamQ+pSB1C3 Colony_1</td></tr>
| |
| - | <tr><td>4</td><td>Pcon+mamQ+pSB1C3 Colony_2</td></tr>
| |
| - | <tr><td>5</td><td>Pcon+mamQ+pSB1C3 Colony_3</td></tr>
| |
| - | <tr><td>6</td><td>Pcon+mamQ+pSB1C3 Colony_4</td></tr>
| |
| - | <tr><td>7</td><td>Pcon+mamQ+pSB1C3 Colony_5</td></tr>
| |
| - | <tr><td>8</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Restriction Enzyme Digestion</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Sample</th><th>DNA/(µl)</th><th>EcoR1/(µl)</th><th>Xba1/(µl)</th><th>Spe1/(µl)</th><th>Pst1/(µl)</th><th>BSA</th><th>Buffur/(µl)</th><th>MilliQ/(µl)</th><th>TOTAL/(µl)</tr>
| |
| - | <tr><td>9/19 Miniprep Pcon+mamL+mamQ+pSB1C3</td><td>34.7</td><td>0</td><td>0</td><td>2</td><td>2</td><td>0.8</td><td>8(B)</td><td>32.5</td><td>80</td></tr>
| |
| - | <tr><td>9/19 Miniprep pSB3T5</td><td>53.3</td><td>2</td><td>0</td><td>2</td><td>0</td><td>0.8</td><td>8(B)</td><td>14</td><td>80</td></tr>
| |
| - | <tr><td>9/19 Miniprep pSB3T5(Nega)</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1(B)</td><td>8</td><td>10</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>PCR</h4>
| |
| - | <span class="author">Tatsui</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Target</th><th>Pcon+mamB+His</th></tr>
| |
| - | <tr><td>KOD Plus/(µl)</td><td>1</td></tr>
| |
| - | <tr><td>10×Buffer/(µl)</td><td>5</td></tr>
| |
| - | <tr><td>2mM dNTPS/(µl)</td><td>5</td></tr>
| |
| - | <tr><td>25mM MgSO4/(µl)</td><td>3</td></tr>
| |
| - | <tr><td>Fw Primer(E+Pcon+X+RBS+mamB)/(µl)</td><td>2</td></tr>
| |
| - | <tr><td>Rv Primer(S+His+mamB)/(µl)</td><td>2</td></tr>
| |
| - | <tr><td>Template/(µl)</td><td>0.5</td></tr>
| |
| - | <tr><td>MilliQ/(µl)</td><td>31.5</td></tr>
| |
| - | <tr><td>Total/(µl)</td><td>50</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <table class="note">
| |
| - | <tr><th>Steps</th><th>Temparature</th><th>Time</th><th>cycle</th></tr>
| |
| - | <tr><td>Pre Denature</td><td>98℃</td><td>2min</td><td>1time</td></tr>
| |
| - | <tr><td>Denature</td><td>98℃</td><td>10sec</td><td>25cycle</td></tr>
| |
| - | <tr><td>Annealing</td><td>55℃</td><td>15sec</td><td>25cycle</td></tr>
| |
| - | <tr><td>Extension</td><td>72℃</td><td>1min</td><td>25cycle</td></tr>
| |
| - | <tr><td>Final elongation</td><td>72℃</td><td>5min</td><td>1time</td></tr>
| |
| - | <tr><td>Final hold</td><td>12℃</td><td>-</td><td>-</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>DNA Purification</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>DNA</th><th>Concentration/(µg/ml)</th><th>260/280</th><th>260/230</th></tr>
| |
| - | <tr><td>9/18 Digestion mamL+pSB1C3(S+P)</td><td>55</td><td>1.66</td><td>1.66</td></tr>
| |
| - | <tr><td>9/19 PCR Pcon+mamB+His</td><td>150</td><td>1.63</td><td>1.52</td></tr>
| |
| - | </table>
| |
| - |
| |
| - | <h4>Restriction Enzyme Digestion</h4>
| |
| - | <span class="author">Honda</span>
| |
| - | <table class="note">
| |
| - | <tr><th>Sample</th><th>DNA/(µl)</th><th>EcoR1/(µl)</th><th>Xba1/(µl)</th><th>Spe1/(µl)</th><th>Pst1/(µl)</th><th>BSA</th><th>Buffur/(µl)</th><th>MilliQ/(µl)</th><th>TOTAL/(µl)</tr>
| |
| - | <tr><td>9/19 Miniprep Pcon+mamL+mamQ+pSB1C3</td><td>34.7</td><td>0</td><td>0</td><td>2</td><td>2</td><td>0.8</td><td>8(B)</td><td>32.5</td><td>80</td></tr>
| |
| - | <tr><td>9/19 Miniprep pSB3T5</td><td>53.3</td><td>2</td><td>0</td><td>2</td><td>0</td><td>0.8</td><td>8(B)</td><td>14</td><td>80</td></tr>
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| - | <tr><td>9/19 Miniprep pSB3T5(Nega)</td><td>1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1(B)</td><td>8</td><td>10</td></tr>
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| - | <h3>9/20</h3>
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| - | <a name="0922" class="kyoto-jump"></a>
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| - | <h3>9/22</h3>
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