Team:Kyoto/Notebook
From 2014.igem.org
(Difference between revisions)
ARIATHEiGEM (Talk | contribs) |
ARIATHEiGEM (Talk | contribs) |
||
Line 146: | Line 146: | ||
</table> | </table> | ||
- | <h4>Electrophoresis | + | <h4>Electrophoresis</h4> |
<span class="author">No name</span> | <span class="author">No name</span> | ||
<table class="note"> | <table class="note"> | ||
Line 193: | Line 193: | ||
<a name="0801" class="kyoto-jump"></a> | <a name="0801" class="kyoto-jump"></a> | ||
<h3>8/1</h3> | <h3>8/1</h3> | ||
- | <h4>Electrophoresis | + | <h4>Electrophoresis</h4> |
<span class="author">Honda, Li and Yamauchi</span> | <span class="author">Honda, Li and Yamauchi</span> | ||
<table class="note"> | <table class="note"> | ||
Line 271: | Line 271: | ||
</table> | </table> | ||
- | <h4>Electrophoresis | + | <h4>Electrophoresis</h4> |
<span class="author">Honda</span> | <span class="author">Honda</span> | ||
<table class="note"> | <table class="note"> | ||
Line 547: | Line 547: | ||
<a name="0905" class="kyoto-jump"></a> | <a name="0905" class="kyoto-jump"></a> | ||
<h3>9/5</h3> | <h3>9/5</h3> | ||
- | <h4>Electrophoresis | + | <h4>Electrophoresis</h4> |
<span class="author">Honda and Matsumoto</span> | <span class="author">Honda and Matsumoto</span> | ||
<table class="note"> | <table class="note"> | ||
- | <tr><th>Lane</th><th> | + | <tr><th>Lane</th><th>Colony PCR ProductsⅠ</td></tr> |
- | <tr><td>1</td><td> | + | <tr><td>1</td><td>1kb ladder</td></tr> |
- | <tr><td>2</td><td> | + | <tr><td>2</td><td>-</td></tr> |
- | <tr><td>3</td><td> | + | <tr><td>3</td><td>-</td></tr> |
- | <tr><td>4</td><td> | + | <tr><td>4</td><td>9/4 pSB1C3(Nega) Colony_1</td></tr> |
- | <tr><td>5</td><td> | + | <tr><td>5</td><td>9/4 pSB1C3(Nega) Colony_2</td></tr> |
- | <tr><td>6</td><td> | + | <tr><td>6</td><td>9/4 pSB1C3(Nega) Colony_3</td></tr> |
+ | <tr><td>7</td><td>9/4 pSB1C3(Nega) Colony_4</td></tr> | ||
+ | <tr><td>8</td><td>9/4 pSB1C3(Nega) Colony_5</td></tr> | ||
+ | <tr><td>9</td><td>9/4 pSB1C3(Nega) Colony_6</td></tr> | ||
+ | <tr><td>10</td><td>9/4 pSB1C3(Nega) Colony_7</td></tr> | ||
+ | <tr><td>11</td><td>9/4 pSB1C3(Nega) Colony_8</td></tr> | ||
+ | <tr><td>12</td><td>9/4 pSB1C3(Nega) Colony_9</td></tr> | ||
+ | <tr><td>13</td><td>9/4 pSB1C3(Nega) Colony_10</td></tr> | ||
+ | <tr><td>14</td><td>9/4 Pcon+pSB2K3 Colony_1</td></tr> | ||
+ | <tr><td>15</td><td>9/4 Pcon+pSB2K3 Colony_2</td></tr> | ||
+ | <tr><td>16</td><td>9/4 Pcon+pSB2K3 Colony_3</td></tr> | ||
+ | <tr><td>17</td><td>1kb ladder</td></tr> | ||
+ | </table> | ||
+ | |||
+ | <table class="note"> | ||
+ | <tr><th>Lane</th><th>Colony PCR ProductsⅡ</td></tr> | ||
+ | <tr><td>1</td><td>1kb ladder</td></tr> | ||
+ | <tr><td>2</td><td>9/4 Pcon+pSB2K3 Colony_4</td></tr> | ||
+ | <tr><td>3</td><td>9/4 Pcon+pSB2K3 Colony_5</td></tr> | ||
+ | <tr><td>4</td><td>9/4 Pcon+pSB2K3 Colony_6</td></tr> | ||
+ | <tr><td>5</td><td>9/4 Pcon+pSB2K3 Colony_7</td></tr> | ||
+ | <tr><td>6</td><td>9/4 Pcon+pSB2K3 Colony_8</td></tr> | ||
+ | <tr><td>7</td><td>9/4 Pcon+pSB2K3 Colony_9</td></tr> | ||
+ | <tr><td>8</td><td>9/4 Pcon+pSB3T5 Colony_10</td></tr> | ||
+ | <tr><td>9</td><td>9/4 pSB3T5(Nega) Colony_1</td></tr> | ||
+ | <tr><td>10</td><td>9/4 pSB3T5(Nega) Colony_2</td></tr> | ||
+ | <tr><td>11</td><td>9/4 Pcon+pSB2K3 Colony_1</td></tr> | ||
+ | <tr><td>12</td><td>9/4 Pcon+pSB2K3 Colony_2</td></tr> | ||
+ | <tr><td>13</td><td>9/4 Pcon+pSB2K3 Colony_3</td></tr> | ||
+ | <tr><td>14</td><td>9/4 Pcon+pSB2K3 Colony_4</td></tr> | ||
+ | <tr><td>15</td><td>9/4 Pcon+pSB2K3 Colony_5</td></tr> | ||
+ | <tr><td>16</td><td>9/4 Pcon+pSB2K3 Colony_6</td></tr> | ||
+ | <tr><td>17</td><td>1kb ladder</td></tr> | ||
+ | </table> | ||
+ | |||
+ | <table class="note"> | ||
+ | <tr><th>Lane</th><th>Colony PCR ProductsⅢ</td></tr> | ||
+ | <tr><td>1</td><td>1kb ladder</td></tr> | ||
+ | <tr><td>2</td><td>9/4 Pcon+pSB2K3 Colony_7</td></tr> | ||
+ | <tr><td>3</td><td>9/4 Pcon+pSB2K3 Colony_8</td></tr> | ||
+ | <tr><td>4</td><td>9/4 Pcon+pSB2K3 Colony_9</td></tr> | ||
+ | <tr><td>5</td><td>9/4 Pcon+pSB2K3 Colony_10</td></tr> | ||
+ | <tr><td>6</td><td>9/4 pSB2K3(Nega) Colony_1</td></tr> | ||
+ | <tr><td>7</td><td>9/4 pSB2K3(Nega) Colony_2</td></tr> | ||
+ | <tr><td>8</td><td>9/4 pSB2K3(Nega) Colony_3</td></tr> | ||
+ | <tr><td>9</td><td>9/4 pSB2K3(Nega) Colony_4</td></tr> | ||
+ | <tr><td>10</td><td>9/4 pSB2K3(Nega) Colony_5</td></tr> | ||
+ | <tr><td>11</td><td>9/4 pSB1C3(Nega) Colony_1</td></tr> | ||
+ | <tr><td>12</td><td>9/4 pSB1C3(Nega) Colony_2</td></tr> | ||
+ | <tr><td>13</td><td>9/4 pSB1C3(Nega) Colony_3</td></tr> | ||
+ | <tr><td>14</td><td>9/4 pSB1C3(Nega) Colony_4</td></tr> | ||
+ | <tr><td>15</td><td>9/4 pSB1C3(Nega) Colony_5</td></tr> | ||
+ | <tr><td>16</td><td>9/4 pSB1C3(Nega) Colony_6</td></tr> | ||
+ | <tr><td>17</td><td>1kb ladder</td></tr> | ||
+ | </table> | ||
+ | |||
+ | <table class="note"> | ||
+ | <tr><th>Lane</th><th>Colony PCR ProductsⅣ</td></tr> | ||
+ | <tr><td>1</td><td>1kb ladder</td></tr> | ||
+ | <tr><td>2</td><td>9/4 pSB1C3(Nega) Colony_7</td></tr> | ||
+ | <tr><td>3</td><td>9/4 pSB1C3(Nega) Colony_8</td></tr> | ||
+ | <tr><td>4</td><td>9/4 pSB1C3(Nega) Colony_9</td></tr> | ||
+ | <tr><td>5</td><td>9/4 pSB1C3(Nega) Colony_10</td></tr> | ||
+ | <tr><td>6</td><td>9/4 Pcon+pSB1C3 Colony_1</td></tr> | ||
+ | <tr><td>7</td><td>9/4 Pcon+pSB1C3 Colony_2</td></tr> | ||
+ | <tr><td>8</td><td>9/4 Pcon+pSB1C3 Colony_3</td></tr> | ||
+ | <tr><td>9</td><td>9/4 Pcon+pSB1C3 Colony_4</td></tr> | ||
+ | <tr><td>10</td><td>9/4 Pcon+pSB1C3 Colony_5</td></tr> | ||
+ | <tr><td>11</td><td>9/4 Pcon+pSB1C3 Colony_6</td></tr> | ||
+ | <tr><td>12</td><td>9/4 Pcon+pSB1C3 Colony_7</td></tr> | ||
+ | <tr><td>13</td><td>9/4 Pcon+pSB1C3 Colony_8</td></tr> | ||
+ | <tr><td>14</td><td>9/4 Pcon+pSB1C3 Colony_9</td></tr> | ||
+ | <tr><td>15</td><td>9/4 Pcon+pSB1C3 Colony_10</td></tr> | ||
+ | <tr><td>16</td><td>8/22 Miniprep BBa_I732095 Colony_3</td></tr> | ||
+ | <tr><td>17</td><td>1kb ladder</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> | </table> | ||
Revision as of 06:30, 26 September 2014
count down
Home | Team | Official Team Profile | Project | Parts | Modeling | Notebook | Safety | Attributions |
Notebook
Calendar
July | ||||||
---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | ||
6 | 7 | 8 | 9 | 10 | 11 | 12 |
13 | 14 | 15 | 16 | 17 | 18 | 19 |
20 | 21 | 22 | 23 | 24 | 25 | 26 |
27 | 28 | 29 | 30 | 31 |
August | ||||||
---|---|---|---|---|---|---|
1 | 2 | |||||
3 | 4 | 5 | 6 | 7 | 8 | 9 |
10 | 11 | 12 | 13 | 14 | 15 | 16 |
17 | 18 | 19 | 20 | 21 | 22 | 23 |
24 | 25 | 26 | 27 | 28 | 29 | 30 |
31 |
September | ||||||
---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | 6 | |
7 | 8 | 9 | 10 | 11 | 12 | 13 |
14 | 15 | 16 | 17 | 18 | 19 | 20 |
21 | 22 | 23 | 24 | 25 | 26 | 27 |
28 | 29 | 30 |
Octorber | ||||||
---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | |||
5 | 6 | 7 | 8 | 9 | 10 | 11 |
12 | 13 | 14 | 15 | 16 | 17 | 18 |
19 | 20 | 21 | 22 | 23 | 24 | 25 |
26 | 27 | 28 | 29 | 30 | 31 |
7/15
transformation
Sample/(µL) | CompetentCells/(µl) | Total/(µl) | Medium | |
---|---|---|---|---|
BBa_J23101(Pcon) | 2 | 20(2/23) | 22 | LB(CP) |
BBa_B0032(RBS) | 2 | 20(2/23) | 22 | LB(CP) |
7/16
transformation
Sample/(µL) | CompetentCells/(µl) | Total/(µl) | Medium | |
---|---|---|---|---|
BBa_B0030(1) | 2 | 20(2/23) | 22 | LB(CP) |
BBa_B0030(2) | 2 | 20(bought) | 22 | LB(CP) |
BBa_B0032(1) | 2 | 20(2/23) | 22 | LB(CP) |
BBa_B0032(2) | 2 | 20(bought) | 22 | LB(CP) |
7/17
Colony PCR
NameⅠ | NameⅡ |
---|---|
7/16 BBa_B0030(1) Colony_1 | 7/16 BBa_B0032(1) Colony_1 |
7/16 BBa_B0030(1) Colony_2 | 7/16 BBa_B0032(1) Colony_2 |
7/16 BBa_B0030(1) Colony_3 | 7/16 BBa_B0032(1) Colony_3 |
7/16 BBa_B0030(2) Colony_1 | 7/16 BBa_B0032(2) Colony_1 |
7/16 BBa_B0030(2) Colony_2 | 7/16 BBa_B0032(2) Colony_2 |
7/16 BBa_B0030(2) Colony_3 | 7/16 BBa_B0032(2) Colony_3 |
Negative Control | - |
- | - |
Liquid Culture
Name | medium |
---|---|
7/16 BBa_B0030(1) Colony_1 | Plusgrow medium(CP) |
7/16 BBa_B0030(1) Colony_2 | Plusgrow medium(CP) |
7/16 BBa_B0030(1) Colony_3 | Plusgrow medium(CP) |
7/16 BBa_B0030(2) Colony_1 | Plusgrow medium(CP) |
7/16 BBa_B0030(2) Colony_2 | Plusgrow medium(CP) |
7/16 BBa_B0030(2) Colony_3 | Plusgrow medium(CP) |
7/16 BBa_B0032(1) Colony_1 | Plusgrow medium(CP) |
7/16 BBa_B0032(1) Colony_2 | Plusgrow medium(CP) |
7/16 BBa_B0032(1) Colony_3 | Plusgrow medium(CP) |
7/16 BBa_B0032(2) Colony_1 | Plusgrow medium(CP) |
7/16 BBa_B0032(2) Colony_2 | Plusgrow medium(CP) |
7/16 BBa_B0032(2) Colony_3 | Plusgrow medium(CP) |
7/18
Miniprep
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
7/16 BBa_B0030(1) Colony_1 | 229 | 1.96 | 1.40 |
7/16 BBa_B0030(1) Colony_2 | 209 | 1.95 | 1.50 |
7/16 BBa_B0030(1) Colony_3 | 108 | 2.12 | 2.12 |
7/16 BBa_B0030(2) Colony_1 | - | - | - |
7/16 BBa_B0030(2) Colony_2 | 174 | 2.02 | 1.56 |
7/16 BBa_B0030(2) Colony_3 | 158 | 2.01 | 1.68 |
7/16 BBa_B0032(1) Colony_1 | 179 | 2.00 | 2.06 |
7/16 BBa_B0032(1) Colony_2 | - | - | - |
7/16 BBa_B0032(1) Colony_3 | 221 | 1.90 | 1.37 |
7/16 BBa_B0032(2) Colony_1 | 225 | 1.89 | 1.37 |
7/16 BBa_B0032(2) Colony_2 | 270 | 1.98 | 1.63 |
7/16 BBa_B0032(2) Colony_3 | 194 | 1.89 | 1.33 |
7/30
Restriction Enzyme Digestion
Sample | DNA/(µl) | EcoR1/(µl) | Xba1/(µl) | Spe1/(µl) | Pst1/(µl) | Buffur/(µl) | BSA/(µl) | MilliQ/(µl) | TOTAL/(µl) |
---|---|---|---|---|---|---|---|---|---|
7/16 BBa_B0030(2) Colony_1 | 1 | 0.3 | 0 | 0 | 0 | 1(H) | 0 | 7.7 | 10 |
7/16 BBa_B0030(2) Colony_1 | 1 | 0 | 0.3 | 0 | 0 | 1(M) | 1 | 6.7 | 10 |
7/16 BBa_B0030(2) Colony_1 | 1 | 0 | 0 | 0.3 | 0 | 1(M) | 0 | 7.7 | 10 |
7/16 BBa_B0030(2) Colony_1 | 1 | 0 | 0 | 0 | 0.3 | 1(H) | 0 | 7.7 | 10 |
7/16 BBa_B0030(2) Colony_1(Nega) | 1 | 0 | 0 | 0 | 0 | 1(M) | 0 | 8 | 10 |
Electrophoresis
Lane | Restriction Enzyme Digestion Products |
---|---|
1 | 7/30 BBa_B0030(2) Colony_1(E CUT) |
2 | 7/30 BBa_B0030(2) Colony_1(X CUT) |
3 | 7/30 BBa_B0030(2) Colony_1(S CUT) |
4 | 7/30 BBa_B0030(2) Colony_1(P CUT) |
5 | 7/30 BBa_B0030(2) Colony_1(Nega) |
6 | 1kbp ladder |
7/31
Transformation
Sample/(µL) | CompetentCells/(µl) | Total/(µl) | Medium | |
---|---|---|---|---|
BBa_E0240(RBS-GFP-DT) | 2 | 20 | 22 | LB(CP) |
J23100(Pcon) | 2 | 20 | 22 | LB(AMP) |
Liquid Culture
Name | medium |
---|---|
7/31 J23100(Pcon) Colony_1 | Plusgrow medium(AMP) |
7/31 J23100(Pcon) Colony_2 | Plusgrow medium(AMP) |
7/31 J23100(Pcon) Colony_3 | Plusgrow medium(AMP) |
7/31 J23100(Pcon) Colony_4 | Plusgrow medium(AMP) |
7/31 J23100(Pcon) Colony_5 | Plusgrow medium(AMP) |
7/31 J23100(Pcon) Colony_6 | Plusgrow medium(AMP) |
Restriction Enzyme Digestion
7/18 RBS Sample | DNA/(µl) | EcoR1/(µl) | Spe1/(µl) | Pst1/(µl) | Xba1/(µl) | Buffur/(µl) | BSA/(µl) | MilliQ/(µl) | TOTAL/(µl) |
---|---|---|---|---|---|---|---|---|---|
BBa_B0032 | 5 | 0.3 | 0 | 0 | 0 | 1(H) | 0 | 3.7 | 10 |
BBa_B0032 | 5 | 0 | 0.3 | 0 | 0 | 1(M) | 0 | 3.7 | 10 |
BBa_B0032 | 5 | 0 | 0 | 0.3 | 0 | 1(H) | 0 | 3.7 | 10 |
BBa_B0032 | 5 | 0 | 0 | 0 | 0.3 | 1(M) | 1 | 2.7 | 10 |
BBa_B0032(Nega) | 5 | 0 | 0 | 0 | 0 | 1(M) | 0 | 4 | 10 |
8/1
Electrophoresis
Lane | Restriction Enzyme Digestion Products |
---|---|
1 | 1kb ladder |
2 | 7/31 BBa_B0032(E CUT) |
3 | 7/31 BBa_B0032(S CUT)Error |
4 | - |
5 | - |
6 | 7/31 BBa_B0032(P CUT) |
7 | 7/31 BBa_B0032(X CUT) |
8 | 7/31 BBa_B0032(Nega) |
PCR
BBa_E0240(RBS-GFP-DT) | Volume |
---|---|
10×buffur | 5μL |
2mMdNTPs | 5μL |
25mM MgSO4 | 2μL |
VF Primer | 1.5μL |
VR Primer | 1.5μL |
template | 0.5μL |
KOD-plus | 1μL |
MilliQ | 33.5μL |
TOTAL | 50μL |
Steps | Temparature | Time | cycle |
---|---|---|---|
Pre Denature | 94℃ | 2min | 1 time |
Denature | 94℃ | 15sec | 30 cycle |
Annealing | 55℃ | 30sec | 30 cycle |
Extension | 68℃ | 1min | 30 cycle |
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
Product | 80 | 1.91 | 1.78 |
8/2
Miniprep
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
7/31 J23100(Pcon) Colony_1 | 145 | 2.11 | 3.09 |
7/31 J23100(Pcon) Colony_2 | - | - | - |
7/31 J23100(Pcon) Colony_3 | 160 | 2.01 | 3.13 |
7/31 J23100(Pcon) Colony_4 | 140 | 1.87 | 2.70 |
7/31 J23100(Pcon) Colony_5 | 105 | 2.39 | 5.90 |
7/31 J23100(Pcon) Colony_6 | 180 | 1.90 | 1.37 |
8/20
Transformation
Sample/(µL) | CompetentCells/(µl) | Total/(µl) | Medium | |
---|---|---|---|---|
7/31 BBa_E0240(RBS-GFD-DT) | 2 | 20 | 22 | LB(CP) |
BBa_I732095 | 2 | 20 | 22 | LB(CP) |
8/21
Colony PCR
NameⅠ |
---|
8/20 BBa_I732095 Colony_1 |
8/20 BBa_I732095 Colony_2 |
8/20 BBa_I732095 Colony_3 |
Negative control |
Electrophoresis
Lane | PCR Product |
---|---|
1 | 1kb ladder |
2 | 8/21 BBa_I732095 Colony_1 |
3 | 8/21 BBa_I732095 Colony_2 |
4 | 8/21 BBa_I732095 Colony_3 |
5 | Negative Control |
Liquid Culture
Name | medium |
---|---|
8/20 BBa_I732095 Colony_1 | Plusgrow medium(CP) |
8/20 BBa_I732095 Colony_2 | Plusgrow medium(CP) |
8/20 BBa_I732095 Colony_3 | Plusgrow medium(CP) |
8/22
Miniprep
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
8/20 BBa_I732095 Colony_1 | 430 | 1.68 | 2.13 |
8/20 BBa_I732095 Colony_2 | 255 | 1.87 | 1.81 |
8/20 BBa_I732095 Colony_3 | 215 | 1.78 | 1.35 |
Restriction Enzyme Digestion
Sample | DNA/(µl) | EcoR1/(µl) | Spe1/(µl) | Buffur/(µl) | MilliQ/(µl) | TOTAL/(µl) |
---|---|---|---|---|---|---|
8/22 BBa_I732095 Colony_1 | 4.7 | 1 | 1 | 3(H) | 20.3 | 30 |
8/22 BBa_I732095 Colony_2 | 7.8 | 1 | 1 | 3(H) | 17.2 | 30 |
8/22 BBa_I732095 Colony_3 | 9.3 | 1 | 1 | 3(H) | 15.7 | 30 |
Transformation
Sample/(µL) | CompetentCells/(µl) | Total/(µl) | Medium | |
---|---|---|---|---|
BBa_J04450(pSB3C5) | 2 | 20 | 22 | LB(CP) |
8/25
Transformation
Sample/(µL) | CompetentCells/(µl) | Total/(µl) | Medium | |
---|---|---|---|---|
BBa_J04450(pSB2K3) | 2 | 20 | 22 | LB(KAN) |
BBa_C0051(pSB1A2) | 2 | 20 | 22 | LB(AMP) |
Liquid Culture
Name | medium |
---|---|
8/22 BBa_J04450(pSB3C5) | Plusgrow medium(CP)*5 |
8/26
Miniprep
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
8/25 BBa_J04450(pSB3C5) | 575 | 1.37 | 1.37 |
Liquid Culture
Name | medium |
---|---|
8/25 BBa_J04450(pSB2K3) | Plusgrow medium(KAN)*5 |
8/25 BBa_J0051(pSB1A2) | Plusgrow medium(AMP)*2 |
8/27
Miniprep
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
8/26 BBa_J04450(pSB2K3) | 1045 | 1.43 | 1.11 |
8/26 BBa_J0051(pSB1A2) | 790 | 1.39 | 0.90 |
Transformation
Sample/(µL) | CompetentCells/(µl) | Total/(µl) | Medium | |
---|---|---|---|---|
BBa_J04450(pSB3T5) | 2 | 20 | 22 | LB(TET) |
8/28
Liquid Culture
Name | medium |
---|---|
8/27 BBa_J04450(pSB3T5) | Plusgrow medium(TET) |
8/29
Miniprep
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
8/28 BBa_J04450(pSB3T5) | 180 | 1.60 | 1.67 |
Transformation
Sample/(µL) | CompetentCells/(µl) | Total/(µl) | Medium | |
---|---|---|---|---|
BBa_J04450(pSB4A5) | 2 | 20 | 22 | LB(AMP) |
9/1
Colony PCR
NameⅠ |
---|
- |
- |
- |
9/1 pSB4A5 Colony_1 |
9/1 pSB4A5 Colony_2 |
Negative Control |
Restriction Enzyme Digestion
Sample | DNA/(µl) | EcoR1/(µl) | Pst1/(µl) | Buffur/(µl) | MilliQ/(µl) | TOTAL/(µl) |
---|---|---|---|---|---|---|
8/28 Miniprep pSB3T5 | 25 | 2 | 2 | 6(H) | 25 | 60 |
Negative control_1 | 3.7 | 0 | 0 | 1(H) | 5.3 | 10 |
8/22 Miniprep I732095 | 14 | 3 | 3 | 9(H) | 61 | 90 |
Negative control_2 | 1.5 | 0 | 0 | 1(H) | 7.5 | 10 |
8/27 Miniprep pSB2K3 | 6 | 3 | 3 | 9(H) | 69 | 90 |
Negative control_3 | 1 | 0 | 0 | 1(H) | 8 | 10 |
8/2 Miniprep Pcon Colony_3 | 20 | 1.5 | 1.5 | 5(H) | 17 | 45 |
Negative control_4 | 4 | 0 | 0 | 1(H) | 5 | 10 |
8/2 Miniprep Pcon Colony_1 | 20 | 1.5 | 1.5 | 5(H) | 17 | 45 |
Negative control_5 | 4 | 0 | 0 | 1(H) | 5 | 10 |
Liquid Culture
Name | medium |
---|---|
8/29 BBa_J04450(pSB4A5) Colony_1 | Plusgrow medium(AMP)*2 |
8/1 Pcon Colony_1 | Plusgrow medium(AMP)*2 |
8/27 pSB3T5 Colony_1 | Plusgrow medium(TET)*2 |
9/2
Miniprep
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
8/29 BBa_J04450(pSB4A5) Colony_1 | 210 | 1.61 | 1.18 |
8/1 Pcon Colony_1 | 215 | 1.79 | 1.87 |
8/27 pSB3T5 Colony_1 | 215 | 1.58 | 0.92 |
Gel Extractions
Lane | Sample | Volume/(µl) |
---|---|---|
1 | 9/1 pSB3T5 Colony_1 (Nega) | 10 |
2 | 9/1 pSB3T5 Colony_1 (E+P)(Confirmation) | 5 |
3 | 9/1 pSB3T5 Colony_1 (E+P)(Extraction) | 55 |
4 | 1kb ladder | 3 |
5 | 9/1 I732095 (Nega) | 10 |
6 | 9/1 I732095 (E+P)(Confirmation) | 5 |
7 | 9/1 I732095 (E+P)(Extraction) | 85 |
8 | 1kb ladder | 3 |
9 | 9/1 pSB2K3 (Nega) | 10 |
10 | 9/1 pSB2K3 (E+P)(Confirmation) | 5 |
11 | 9/1 pSB2K3 (E+P)(Extraction) | 85 |
Lane | Sample | Volume/(µl) |
---|---|---|
1 | 9/1 Pcon Colony_3 (E+P)(Confirmation) | 5 |
2 | 9/1 Pcon Colony_3 (E+P)(Extraction) | 20 |
3 | 9/1 Pcon Colony_3 (E+P)(Extraction) | 20 |
4 | 9/1 Pcon Colony_3 (Nega) | 10 |
5 | 1kb ladder | 3 |
6 | 9/1 Pcon Colony_1 (E+P)(Confirmation) | 5 |
7 | 9/1 Pcon Colony_1 (E+P)(Extraction) | 20 |
8 | 9/1 Pcon Colony_1 (E+P)(Extraction) | 20 |
9 | 9/1 Pcon Colony_1 (Nega) | 10 |
9/3
Concentration Measurement
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
pSB2K3 | 40 | 1.48 | 0.10 |
Pcon Colony_1 | 30 | 1.76 | 0.62 |
pSB3T5 | 50 | 2.35 | 0.50 |
pSB1C3 | 65 | 2.14 | 0.10 |
Pcon Colony_3 | 45 | 1.56 | 0.58 |
Ligation
Insert/(µL) | Vector/(µL) | T4 Ligase/(µl) | 2*Buffer/(µl) | MilliQ/(µl) | Total/(µl) | ||
---|---|---|---|---|---|---|---|
Pcon | 2.1 | pSB1C3 | 1.5 | 1 | 5 | 0.4 | 10 |
-(Negative Control) | 0 | pSB1C3 | 1.5 | 1 | 5 | 2.5 | 10 |
Pcon | 1.3 | pSB2K3 | 2.5 | 1 | 5 | 0.2 | 10 |
-(Negative Control) | 0 | pSB2K3 | 2.5 | 1 | 5 | 1.5 | 10 |
Pcon | 10 | pSB3T5 | 2.3 | 1 | 5 | 0.7 | 10 |
-(Negative Control) | 0 | pSB3T5 | 2.3 | 1 | 5 | 1.7 | 10 |
Transformation
Sample/(µL) | CompetentCells/(µl) | Total/(µl) | Medium | |
---|---|---|---|---|
Pcon+pSB1C3 | 3 | 15 | 18 | LB(CP) |
pSB1C3(Negative Control) | 8 | 10 | 18 | LB(CP) |
Pcon+pSB2K3 | 3 | 15 | 18 | LB(KAN) |
pSB2K3(Negative Control) | 8 | 10 | 18 | LB(KAN) |
Pcon+pSB3T5 | 3 | 15 | 18 | LB(TET) |
pSB3T5(Negative Control) | 2 | 10 | 12 | LB(TET) |
9/4
Colony PCR
NameⅠ | NameⅡ | NameⅢ | NameⅣ |
---|---|---|---|
- | 9/3 pSB1C3(Nega) Colony_7 | 9/3 Pcon+pSB3T5 Colony_5 | 9/3 Pcon+pSB2K3 Colony_1 |
- | 9/3 pSB1C3(Nega) Colony_8 | 9/3 Pcon+pSB3T5 Colony_6 | 9/3 Pcon+pSB2K3 Colony_2 |
9/3 pSB1C3(Nega) Colony_1 | 9/3 Pcon+pSB3T5 Colony_1 | 9/3 Pcon+pSB3T5 Colony_7 | 9/3 Pcon+pSB2K3 Colony_3 |
9/3 pSB1C3(Nega) Colony_2 | 9/3 Pcon+pSB3T5 Colony_2 | 9/3 Pcon+pSB3T5 Colony_8 | 9/3 Pcon+pSB2K3 Colony_4 |
9/3 pSB1C3(Nega) Colony_3 | 9/3 Pcon+pSB3T5 Colony_3 | 9/3 Pcon+pSB3T5 Colony_9 | 9/3 Pcon+pSB2K3 Colony_5 |
9/3 pSB1C3(Nega) Colony_4 | 9/3 Pcon+pSB3T5 Colony_4 | 9/3 Pcon+pSB3T5 Colony_10 | 9/3 Pcon+pSB2K3 Colony_6 |
9/3 pSB1C3(Nega) Colony_5 | 9/3 pSB1C3(Nega) Colony_9 | 9/3 pSB3T5(Nega) Colony_1 | 9/3 Pcon+pSB2K3 Colony_7 |
9/3 pSB1C3(Nega) Colony_6 | 9/3 pSB1C3(Nega) Colony_10 | 9/3 pSB3T5(Nega) Colony_2 | 9/3 Pcon+pSB2K3 Colony_8 |
NameⅤ | NameⅥ | NameⅦ | NameⅧ |
---|---|---|---|
9/3 Pcon+pSB2K3 Colony_9 | 9/3 Pcon+pSB1C3 Colony_2 | 9/3 Pcon+pSB1C3 Colony_10 | 9/3 pSB1C3(Nega) Colony_8 |
9/3 Pcon+pSB2K3 Colony_10 | 9/3 Pcon+pSB1C3 Colony_3 | 9/3 pSB1C3(Nega) Colony_1 | 9/3 pSB1C3(Nega) Colony_9 |
9/3 pSB2K3(Nega) Colony_1 | 9/3 Pcon+pSB1C3 Colony_4 | 9/3 pSB1C3(Nega) Colony_2 | 9/3 pSB1C3(Nega) Colony_10 |
9/3 pSB2K3(Nega) Colony_2 | 9/3 Pcon+pSB1C3 Colony_5 | 9/3 pSB1C3(Nega) Colony_3 | 9/3 pSB1C3(Nega) Colony_11 |
9/3 pSB2K3(Nega) Colony_3 | 9/3 Pcon+pSB1C3 Colony_6 | 9/3 pSB1C3(Nega) Colony_4 | - |
9/3 pSB2K3(Nega) Colony_4 | 9/3 Pcon+pSB1C3 Colony_7 | 9/3 pSB1C3(Nega) Colony_5 | - |
9/3 pSB2K3(Nega) Colony_5 | 9/3 Pcon+pSB1C3 Colony_8 | 9/3 pSB1C3(Nega) Colony_6 | - |
9/3 Pcon+pSB1C3 Colony_1 | 9/3 Pcon+pSB1C3 Colony_9 | 9/3 pSB1C3(Nega) Colony_7 | - |
NameⅨ | NameⅩ |
---|---|
9/3 Pcon+pSB1C3 Colony_11 | 8/22 Miniprep I732095 Colony_3 |
Negative Control | - |
9/5
Electrophoresis
Lane | Colony PCR ProductsⅠ |
---|---|
1 | 1kb ladder |
2 | - |
3 | - |
4 | 9/4 pSB1C3(Nega) Colony_1 |
5 | 9/4 pSB1C3(Nega) Colony_2 |
6 | 9/4 pSB1C3(Nega) Colony_3 |
7 | 9/4 pSB1C3(Nega) Colony_4 |
8 | 9/4 pSB1C3(Nega) Colony_5 |
9 | 9/4 pSB1C3(Nega) Colony_6 |
10 | 9/4 pSB1C3(Nega) Colony_7 |
11 | 9/4 pSB1C3(Nega) Colony_8 |
12 | 9/4 pSB1C3(Nega) Colony_9 |
13 | 9/4 pSB1C3(Nega) Colony_10 |
14 | 9/4 Pcon+pSB2K3 Colony_1 |
15 | 9/4 Pcon+pSB2K3 Colony_2 |
16 | 9/4 Pcon+pSB2K3 Colony_3 |
17 | 1kb ladder |
Lane | Colony PCR ProductsⅡ |
---|---|
1 | 1kb ladder |
2 | 9/4 Pcon+pSB2K3 Colony_4 |
3 | 9/4 Pcon+pSB2K3 Colony_5 |
4 | 9/4 Pcon+pSB2K3 Colony_6 |
5 | 9/4 Pcon+pSB2K3 Colony_7 |
6 | 9/4 Pcon+pSB2K3 Colony_8 |
7 | 9/4 Pcon+pSB2K3 Colony_9 |
8 | 9/4 Pcon+pSB3T5 Colony_10 |
9 | 9/4 pSB3T5(Nega) Colony_1 |
10 | 9/4 pSB3T5(Nega) Colony_2 |
11 | 9/4 Pcon+pSB2K3 Colony_1 |
12 | 9/4 Pcon+pSB2K3 Colony_2 |
13 | 9/4 Pcon+pSB2K3 Colony_3 |
14 | 9/4 Pcon+pSB2K3 Colony_4 |
15 | 9/4 Pcon+pSB2K3 Colony_5 |
16 | 9/4 Pcon+pSB2K3 Colony_6 |
17 | 1kb ladder |
Lane | Colony PCR ProductsⅢ |
---|---|
1 | 1kb ladder |
2 | 9/4 Pcon+pSB2K3 Colony_7 |
3 | 9/4 Pcon+pSB2K3 Colony_8 |
4 | 9/4 Pcon+pSB2K3 Colony_9 |
5 | 9/4 Pcon+pSB2K3 Colony_10 |
6 | 9/4 pSB2K3(Nega) Colony_1 |
7 | 9/4 pSB2K3(Nega) Colony_2 |
8 | 9/4 pSB2K3(Nega) Colony_3 |
9 | 9/4 pSB2K3(Nega) Colony_4 |
10 | 9/4 pSB2K3(Nega) Colony_5 |
11 | 9/4 pSB1C3(Nega) Colony_1 |
12 | 9/4 pSB1C3(Nega) Colony_2 |
13 | 9/4 pSB1C3(Nega) Colony_3 |
14 | 9/4 pSB1C3(Nega) Colony_4 |
15 | 9/4 pSB1C3(Nega) Colony_5 |
16 | 9/4 pSB1C3(Nega) Colony_6 |
17 | 1kb ladder |
Lane | Colony PCR ProductsⅣ |
---|---|
1 | 1kb ladder |
2 | 9/4 pSB1C3(Nega) Colony_7 |
3 | 9/4 pSB1C3(Nega) Colony_8 |
4 | 9/4 pSB1C3(Nega) Colony_9 |
5 | 9/4 pSB1C3(Nega) Colony_10 |
6 | 9/4 Pcon+pSB1C3 Colony_1 |
7 | 9/4 Pcon+pSB1C3 Colony_2 |
8 | 9/4 Pcon+pSB1C3 Colony_3 |
9 | 9/4 Pcon+pSB1C3 Colony_4 |
10 | 9/4 Pcon+pSB1C3 Colony_5 |
11 | 9/4 Pcon+pSB1C3 Colony_6 |
12 | 9/4 Pcon+pSB1C3 Colony_7 |
13 | 9/4 Pcon+pSB1C3 Colony_8 |
14 | 9/4 Pcon+pSB1C3 Colony_9 |
15 | 9/4 Pcon+pSB1C3 Colony_10 |
16 | 8/22 Miniprep BBa_I732095 Colony_3 |
17 | 1kb ladder |
Lane | Colony PCR ProductsⅤ |
---|---|
1 | 1kb ladder |
2 | 9/4 Pcon+pSB1C3 Colony_11 |
3 | 9/4 Negative Control |
4 | 8/22 Miniprep BBa_I732095 Colony_3 |
5 | - |
6 | - |
7 | - |
8 | - |