From 2014.igem.org
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| <p>Objective: test backbone shuffled antitracr bacteria</p> | | <p>Objective: test backbone shuffled antitracr bacteria</p> |
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- | <ul><li>Started colonies from plates and diluted in to +/- aTc this morning. Let shake for 5(?) hours before collecting 500uL and spin/resusp in PBS for flow.</li><li>Took data to flowjo and plotted histograms below. This is ungated fluorescence</li><li></li><li><span style="overflow: hidden; display: inline-block; margin: 0.00px 0.00px; border: 0.00px solid #000000; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px); width: 624.00px; height: 482.67px;"><img alt="" src="images/image44.jpg" style="width: 624.00px; height: 482.67px; margin-left: 0.00px; margin-top: 0.00px; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px);" title=""></li><li>table with mean GFPAs</li></ul><a href="#" name="096a26566c8f532b5e11f6f110d57e6edec006da"></a><a href="#" name="3"></a><table cellpadding="0" cellspacing="0"><tbody><tr><td colspan="1" rowspan="1"><p></p></td><td colspan="1" rowspan="1"><p></p></td><td colspan="1" rowspan="1"><p></p></td><td colspan="1" rowspan="1"><p>ungated GFPA</p></td><td colspan="1" rowspan="1"><p>+/- aTc</p></td></tr><tr><td colspan="1" rowspan="1"><p>1: if142014-09-04.001.fcs</p></td><td colspan="1" rowspan="1"><p>AC-1</p></td><td colspan="1" rowspan="1"><p>+ aTc</p></td><td colspan="1" rowspan="1"><p>0.402</p></td><td colspan="1" rowspan="1"><p>1.185840708</p></td></tr><tr><td colspan="1" rowspan="1"><p>2: if142014-09-04.002.fcs</p></td><td colspan="1" rowspan="1"><p>AC-2</p></td><td colspan="1" rowspan="1"><p>+ aTc</p></td><td colspan="1" rowspan="1"><p>0.455</p></td><td colspan="1" rowspan="1"><p>1.197368421</p></td></tr><tr><td colspan="1" rowspan="1"><p>3: if142014-09-04.003.fcs</p></td><td colspan="1" rowspan="1"><p>G-1</p></td><td colspan="1" rowspan="1"><p>+ aTc</p></td><td colspan="1" rowspan="1"><p>14.9</p></td><td colspan="1" rowspan="1"><p>0.949044586</p></td></tr><tr><td colspan="1" rowspan="1"><p>4: if142014-09-04.004.fcs</p></td><td colspan="1" rowspan="1"><p>G-2</p></td><td colspan="1" rowspan="1"><p>+ aTc</p></td><td colspan="1" rowspan="1"><p>11.7</p></td><td colspan="1" rowspan="1"><p>0.573529412</p></td></tr><tr><td colspan="1" rowspan="1"><p>5: if142014-09-04.005.fcs</p></td><td colspan="1" rowspan="1"><p>G-3</p></td><td colspan="1" rowspan="1"><p>+ aTc</p></td><td colspan="1" rowspan="1"><p>8.91</p></td><td colspan="1" rowspan="1"><p>0.530357143</p></td></tr><tr><td colspan="1" rowspan="1"><p>6: if142014-09-04.006.fcs</p></td><td colspan="1" rowspan="1"><p>G-4</p></td><td colspan="1" rowspan="1"><p>+ aTc</p></td><td colspan="1" rowspan="1"><p>5.92</p></td><td colspan="1" rowspan="1"><p>0.34619883</p></td></tr><tr><td colspan="1" rowspan="1"><p>7: if142014-09-04.007.fcs</p></td><td colspan="1" rowspan="1"><p>C-1</p></td><td colspan="1" rowspan="1"><p>+ aTc</p></td><td colspan="1" rowspan="1"><p>186</p></td><td colspan="1" rowspan="1"><p>1.016393443</p></td></tr><tr><td colspan="1" rowspan="1"><p>8: if142014-09-04.008.fcs</p></td><td colspan="1" rowspan="1"><p>C-2</p></td><td colspan="1" rowspan="1"><p>+ aTc</p></td><td colspan="1" rowspan="1"><p>224</p></td><td colspan="1" rowspan="1"><p>1.230769231</p></td></tr><tr><td colspan="1" rowspan="1"><p>9: if142014-09-04.009.fcs</p></td><td colspan="1" rowspan="1"><p>C-3</p></td><td colspan="1" rowspan="1"><p>+ aTc</p></td><td colspan="1" rowspan="1"><p>214</p></td><td colspan="1" rowspan="1"><p>0.922413793</p></td></tr><tr><td colspan="1" rowspan="1"><p>10: if142014-09-04.010.fcs</p></td><td colspan="1" rowspan="1"><p>C-4</p></td><td colspan="1" rowspan="1"><p>+ aTc</p></td><td colspan="1" rowspan="1"><p>222</p></td><td colspan="1" rowspan="1"><p>1.193548387</p></td></tr><tr><td colspan="1" rowspan="1"><p>11: if142014-09-04.011.fcs</p></td><td colspan="1" rowspan="1"><p>AC-1</p></td><td colspan="1" rowspan="1"><p>- aTc</p></td><td colspan="1" rowspan="1"><p>0.339</p></td><td colspan="1" rowspan="1"><p></p></td></tr><tr><td colspan="1" rowspan="1"><p>12: if142014-09-04.012.fcs</p></td><td colspan="1" rowspan="1"><p>AC-2</p></td><td colspan="1" rowspan="1"><p>- aTc</p></td><td colspan="1" rowspan="1"><p>0.38</p></td><td colspan="1" rowspan="1"><p></p></td></tr><tr><td colspan="1" rowspan="1"><p>13: if142014-09-04.013.fcs</p></td><td colspan="1" rowspan="1"><p>G-1</p></td><td colspan="1" rowspan="1"><p>- aTc</p></td><td colspan="1" rowspan="1"><p>15.7</p></td><td colspan="1" rowspan="1"><p></p></td></tr><tr><td colspan="1" rowspan="1"><p>14: if142014-09-04.014.fcs</p></td><td colspan="1" rowspan="1"><p>G-2</p></td><td colspan="1" rowspan="1"><p>- aTc</p></td><td colspan="1" rowspan="1"><p>20.4</p></td><td colspan="1" rowspan="1"><p></p></td></tr><tr><td colspan="1" rowspan="1"><p>15: if142014-09-04.015.fcs</p></td><td colspan="1" rowspan="1"><p>G-3</p></td><td colspan="1" rowspan="1"><p>- aTc</p></td><td colspan="1" rowspan="1"><p>16.8</p></td><td colspan="1" rowspan="1"><p></p></td></tr><tr><td colspan="1" rowspan="1"><p>16: if142014-09-04.016.fcs</p></td><td colspan="1" rowspan="1"><p>G-4</p></td><td colspan="1" rowspan="1"><p>- aTc</p></td><td colspan="1" rowspan="1"><p>17.1</p></td><td colspan="1" rowspan="1"><p></p></td></tr><tr><td colspan="1" rowspan="1"><p>17: if142014-09-04.017.fcs</p></td><td colspan="1" rowspan="1"><p>C-1</p></td><td colspan="1" rowspan="1"><p>- aTc</p></td><td colspan="1" rowspan="1"><p>183</p></td><td colspan="1" rowspan="1"><p></p></td></tr><tr><td colspan="1" rowspan="1"><p>18: if142014-09-04.018.fcs</p></td><td colspan="1" rowspan="1"><p>C-2</p></td><td colspan="1" rowspan="1"><p>- aTc</p></td><td colspan="1" rowspan="1"><p>182</p></td><td colspan="1" rowspan="1"><p></p></td></tr><tr><td colspan="1" rowspan="1"><p>19: if142014-09-04.019.fcs</p></td><td colspan="1" rowspan="1"><p>C-3</p></td><td colspan="1" rowspan="1"><p>- aTc</p></td><td colspan="1" rowspan="1"><p>232</p></td><td colspan="1" rowspan="1"><p></p></td></tr><tr><td colspan="1" rowspan="1"><p>20: if142014-09-04.020.fcs</p></td><td colspan="1" rowspan="1"><p>C-4</p></td><td colspan="1" rowspan="1"><p>- aTc</p></td><td colspan="1" rowspan="1"><p>186</p></td><td colspan="1" rowspan="1"><p></p></td></tr></tbody></table><ul><li>aTc treatment seems to be moving fluor in the opposite direction of what we’d expect of antitracr was doing its job. The degree is inconsistent, reflow is called for. </li></ul><p></p><p></p><p></p><p></p><hr style="page-break-before:always;display:none;"><p></p><hr style="page-break-before:always;display:none;"><p></p><p></p><h2><a name="h.ohhk3mdbkmcm"></a>9/8/14</h2><p>Objective: retest backbone shuffled antitracr bacteria</p><ul><li>Yesterday picked 4 new colonies from plates and 4 from 9/4/14 frozen stocks and grew in LB + antibiotics overnight. Diluted in to +/- aTc this morning and let shake for 5(?) hours before collecting 500uL and spin/resusp in PBS</li><li>G1-4 are old frozen coli with K608012, pdCas9-GFP1, and 1A2-R0040-antitracr, C1-4 are old frozen with pdCas9. clones 5-8 are new from plates. AC are no-fluor (1A2 and pdCas9) but they were diluted in to triple antibiotic medium instead of double so they did not grow and are not reliable indicators of bg fluorescence on this run.</li><li>After flow, isolated singlets with flowjo (smalls) or left alone (bugs) and got gfpa unmodified or normalized by fsca or ssca. </li><li></li></ul><a href="#" name="777f62c089f7173b14ec6d88236db6c3e53afff8"></a><a href="#" name="4"></a><table cellpadding="0" cellspacing="0"<><tbody><tr><td colspan="1" rowspan="1"><p></p></td><td colspan="1" rowspan="1"><p></p></td><td colspan="1" rowspan="1"><p></p></td><td colspan="1" rowspan="1"><p></p></td><td colspan="1" rowspan="1"><p>smalls</p></td><td colspan="1" rowspan="1"><p></p></td><td colspan="1" rowspan="1"><p></p></td><td colspan="1" rowspan="1"><p>bugs</p></td><td colspan="1" rowspan="1"><p></p></td></tr><tr><td colspan="1" rowspan="1"><p></p></td><td colspan="1" rowspan="1"><p></p></td><td colspan="1" rowspan="1"><p>aTc</p></td><td colspan="1" rowspan="1"><p>gfpa</p></td><td colspan="1" rowspan="1"><p>gfpa/fsca*1000</p></td><td colspan="1" rowspan="1"><p>gfpa/ssca*1000</p></td><td colspan="1" rowspan="1"><p>gfpa</p></td><td colspan="1" rowspan="1"><p>gfpa/fsca*1000</p></td><td colspan="1" rowspan="1"><p>gfpa/ssca*1000</p></td></tr><tr><td colspan="1" rowspan="1"><p>1: if142014-09-08.001.fcs</p></td><td colspan="1" rowspan="1"><p>G1</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>5.25</p></td><td colspan="1" rowspan="1"><p>243</p></td><td colspan="1" rowspan="1"><p>112</p></td><td colspan="1" rowspan="1"><p>7.79</p></td><td colspan="1" rowspan="1"><p>247</p></td><td colspan="1" rowspan="1"><p>114</p></td></tr><tr><td colspan="1" rowspan="1"><p>2: if142014-09-08.002.fcs</p></td><td colspan="1" rowspan="1"><p>G2</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>4.41</p></td><td colspan="1" rowspan="1"><p>226</p></td><td colspan="1" rowspan="1"><p>78.1</p></td><td colspan="1" rowspan="1"><p>7.12</p></td><td colspan="1" rowspan="1"><p>228</p></td><td colspan="1" rowspan="1"><p>76.2</p></td></tr><tr><td colspan="1" rowspan="1"><p>3: if142014-09-08.003.fcs</p></td><td colspan="1" rowspan="1"><p>G3</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>4.66</p></td><td colspan="1" rowspan="1"><p>231</p></td><td colspan="1" rowspan="1"><p>84.8</p></td><td colspan="1" rowspan="1"><p>7.8</p></td><td colspan="1" rowspan="1"><p>231</p></td><td colspan="1" rowspan="1"><p>83.4</p></td></tr><tr><td colspan="1" rowspan="1"><p>4: if142014-09-08.004.fcs</p></td><td colspan="1" rowspan="1"><p>G4</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>3.86</p></td><td colspan="1" rowspan="1"><p>191</p></td><td colspan="1" rowspan="1"><p>70.1</p></td><td colspan="1" rowspan="1"><p>5.99</p></td><td colspan="1" rowspan="1"><p>192</p></td><td colspan="1" rowspan="1"><p>67.9</p></td></tr><tr><td colspan="1" rowspan="1"><p>5: if142014-09-08.005.fcs</p></td><td colspan="1" rowspan="1"><p>G1</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>6.81</p></td><td colspan="1" rowspan="1"><p>311</p></td><td colspan="1" rowspan="1"><p>145</p></td><td colspan="1" rowspan="1"><p>8.56</p></td><td colspan="1" rowspan="1"><p>310</p></td><td colspan="1" rowspan="1"><p>143</p></td></tr><tr><td colspan="1" rowspan="1"><p>6: if142014-09-08.006.fcs</p></td><td colspan="1" rowspan="1"><p>G2</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>6.37</p></td><td colspan="1" rowspan="1"><p>337</p></td><td colspan="1" rowspan="1"><p>153</p></td><td colspan="1" rowspan="1"><p>7.25</p></td><td colspan="1" rowspan="1"><p>323</p></td><td colspan="1" rowspan="1"><p>145</p></td></tr><tr><td colspan="1" rowspan="1"><p>7: if142014-09-08.007.fcs</p></td><td colspan="1" rowspan="1"><p>G3</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>6.5</p></td><td colspan="1" rowspan="1"><p>318</p></td><td colspan="1" rowspan="1"><p>143</p></td><td colspan="1" rowspan="1"><p>8.24</p></td><td colspan="1" rowspan="1"><p>305</p></td><td colspan="1" rowspan="1"><p>136</p></td></tr><tr><td colspan="1" rowspan="1"><p>8: if142014-09-08.008.fcs</p></td><td colspan="1" rowspan="1"><p>G4</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>6.14</p></td><td colspan="1" rowspan="1"><p>298</p></td><td colspan="1" rowspan="1"><p>140</p></td><td colspan="1" rowspan="1"><p>7.66</p></td><td colspan="1" rowspan="1"><p>288</p></td><td colspan="1" rowspan="1"><p>131</p></td></tr><tr><td colspan="1" rowspan="1"><p>9: if142014-09-08.009.fcs</p></td><td colspan="1" rowspan="1"><p>C1</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>88.9</p></td><td colspan="1" rowspan="1"><p>4071</p></td><td colspan="1" rowspan="1"><p>1777</p></td><td colspan="1" rowspan="1"><p>117</p></td><td colspan="1" rowspan="1"><p>3978</p></td><td colspan="1" rowspan="1"><p>1706</p></td></tr><tr><td colspan="1" rowspan="1"><p>10: if142014-09-08.010.fcs</p></td><td colspan="1" rowspan="1"><p>C2</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>83.5</p></td><td colspan="1" rowspan="1"><p>4073</p></td><td colspan="1" rowspan="1"><p>1547</p></td><td colspan="1" rowspan="1"><p>120</p></td><td colspan="1" rowspan="1"><p>3969</p></td><td colspan="1" rowspan="1"><p>1422</p></td></tr><tr><td colspan="1" rowspan="1"><p>11: if142014-09-08.011.fcs</p></td><td colspan="1" rowspan="1"><p>C3</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>81.7</p></td><td colspan="1" rowspan="1"><p>3957</p></td><td colspan="1" rowspan="1"><p>1551</p></td><td colspan="1" rowspan="1"><p>115</p></td><td colspan="1" rowspan="1"><p>3840</p></td><td colspan="1" rowspan="1"><p>1437</p></td></tr><tr><td colspan="1" rowspan="1"><p>12: if142014-09-08.012.fcs</p></td><td colspan="1" rowspan="1"><p>C4</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>77.2</p></td><td colspan="1" rowspan="1"><p>3915</p></td><td colspan="1" rowspan="1"><p>1577</p></td><td colspan="1" rowspan="1"><p>95.7</p></td><td colspan="1" rowspan="1"><p>3883</p></td><td colspan="1" rowspan="1"><p>1503</p></td></tr><tr><td colspan="1" rowspan="1"><p>13: if142014-09-08.013.fcs</p></td><td colspan="1" rowspan="1"><p>C1</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>77.7</p></td><td colspan="1" rowspan="1"><p>3686</p></td><td colspan="1" rowspan="1"><p>1471</p></td><td colspan="1" rowspan="1"><p>100</p></td><td colspan="1" rowspan="1"><p>3545</p></td><td colspan="1" rowspan="1"><p>1411</p></td></tr><tr><td colspan="1" rowspan="1"><p>14: if142014-09-08.014.fcs</p></td><td colspan="1" rowspan="1"><p>C2</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>64.5</p></td><td colspan="1" rowspan="1"><p>3093</p></td><td colspan="1" rowspan="1"><p>1445</p></td><td colspan="1" rowspan="1"><p>88.3</p></td><td colspan="1" rowspan="1"><p>2962</p></td><td colspan="1" rowspan="1"><p>1373</p></td></tr><tr><td colspan="1" rowspan="1"><p>15: if142014-09-08.015.fcs</p></td><td colspan="1" rowspan="1"><p>C3</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>80.9</p></td><td colspan="1" rowspan="1"><p>3971</p></td><td colspan="1" rowspan="1"><p>1541</p></td><td colspan="1" rowspan="1"><p>106</p></td><td colspan="1" rowspan="1"><p>3771</p></td><td colspan="1" rowspan="1"><p>1432</p></td></tr><tr><td colspan="1" rowspan="1"><p>16: if142014-09-08.016.fcs</p></td><td colspan="1" rowspan="1"><p>C4</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>58.9</p></td><td colspan="1" rowspan="1"><p>2899</p></td><td colspan="1" rowspan="1"><p>1299</p></td><td colspan="1" rowspan="1"><p>79.7</p></td><td colspan="1" rowspan="1"><p>2727</p></td><td colspan="1" rowspan="1"><p>1220</p></td></tr><tr><td colspan="1" rowspan="1"><p>17: if142014-09-08.017.fcs</p></td><td colspan="1" rowspan="1"><p>G5</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>4.1</p></td><td colspan="1" rowspan="1"><p>210</p></td><td colspan="1" rowspan="1"><p>79.1</p></td><td colspan="1" rowspan="1"><p>5.4</p></td><td colspan="1" rowspan="1"><p>209</p></td><td colspan="1" rowspan="1"><p>76.2</p></td></tr><tr><td colspan="1" rowspan="1"><p>18: if142014-09-08.018.fcs</p></td><td colspan="1" rowspan="1"><p>G6</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>4.9</p></td><td colspan="1" rowspan="1"><p>225</p></td><td colspan="1" rowspan="1"><p>98.2</p></td><td colspan="1" rowspan="1"><p>7.07</p></td><td colspan="1" rowspan="1"><p>226</p></td><td colspan="1" rowspan="1"><p>97.6</p></td></tr><tr><td colspan="1" rowspan="1"><p>19: if142014-09-08.019.fcs</p></td><td colspan="1" rowspan="1"><p>G7</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>4.42</p></td><td colspan="1" rowspan="1"><p>220</p></td><td colspan="1" rowspan="1"><p>84.7</p></td><td colspan="1" rowspan="1"><p>5.97</p></td><td colspan="1" rowspan="1"><p>218</p></td><td colspan="1" rowspan="1"><p>81.7</p></td></tr><tr><td colspan="1" rowspan="1"><p>20: if142014-09-08.020.fcs</p></td><td colspan="1" rowspan="1"><p>G8</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>4.19</p></td><td colspan="1" rowspan="1"><p>211</p></td><td colspan="1" rowspan="1"><p>81.3</p></td><td colspan="1" rowspan="1"><p>5.68</p></td><td colspan="1" rowspan="1"><p>213</p></td><td colspan="1" rowspan="1"><p>79.5</p></td></tr><tr><td colspan="1" rowspan="1"><p>21: if142014-09-08.021.fcs</p></td><td colspan="1" rowspan="1"><p>G5</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>9.73</p></td><td colspan="1" rowspan="1"><p>477</p></td><td colspan="1" rowspan="1"><p>201</p></td><td colspan="1" rowspan="1"><p>12.5</p></td><td colspan="1" rowspan="1"><p>455</p></td><td colspan="1" rowspan="1"><p>190</p></td></tr><tr><td colspan="1" rowspan="1"><p>22: if142014-09-08.022.fcs</p></td><td colspan="1" rowspan="1"><p>G6</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>9.68</p></td><td colspan="1" rowspan="1"><p>450</p></td><td colspan="1" rowspan="1"><p>197</p></td><td colspan="1" rowspan="1"><p>12.9</p></td><td colspan="1" rowspan="1"><p>449</p></td><td colspan="1" rowspan="1"><p>196</p></td></tr><tr><td colspan="1" rowspan="1"><p>23: if142014-09-08.023.fcs</p></td><td colspan="1" rowspan="1"><p>G7</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>10.1</p></td><td colspan="1" rowspan="1"><p>482</p></td><td colspan="1" rowspan="1"><p>212</p></td><td colspan="1" rowspan="1"><p>13.3</p></td><td colspan="1" rowspan="1"><p>464</p></td><td colspan="1" rowspan="1"><p>198</p></td></tr><tr><td colspan="1" rowspan="1"><p>24: if142014-09-08.024.fcs</p></td><td colspan="1" rowspan="1"><p>G8</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>10.1</p></td><td colspan="1" rowspan="1"><p>489</p></td><td colspan="1" rowspan="1"><p>216</p></td><td colspan="1" rowspan="1"><p>13.3</p></td><td colspan="1" rowspan="1"><p>473</p></td><td colspan="1" rowspan="1"><p>204</p></td></tr><tr><td colspan="1" rowspan="1"><p>25: if142014-09-08.025.fcs</p></td><td colspan="1" rowspan="1"><p>C5</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>79.6</p></td><td colspan="1" rowspan="1"><p>3958</p></td><td colspan="1" rowspan="1"><p>1486</p></td><td colspan="1" rowspan="1"><p>113</p></td><td colspan="1" rowspan="1"><p>3813</p></td><td colspan="1" rowspan="1"><p>1374</p></td></tr><tr><td colspan="1" rowspan="1"><p>26: if142014-09-08.026.fcs</p></td><td colspan="1" rowspan="1"><p>C6</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>80.8</p></td><td colspan="1" rowspan="1"><p>4045</p></td><td colspan="1" rowspan="1"><p>1514</p></td><td colspan="1" rowspan="1"><p>113</p></td><td colspan="1" rowspan="1"><p>4043</p></td><td colspan="1" rowspan="1"><p>1425</p></td></tr><tr><td colspan="1" rowspan="1"><p>27: if142014-09-08.027.fcs</p></td><td colspan="1" rowspan="1"><p>C7</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>80.7</p></td><td colspan="1" rowspan="1"><p>4053</p></td><td colspan="1" rowspan="1"><p>1491</p></td><td colspan="1" rowspan="1"><p>116</p></td><td colspan="1" rowspan="1"><p>3921</p></td><td colspan="1" rowspan="1"><p>1375</p></td></tr><tr><td colspan="1" rowspan="1"><p>28: if142014-09-08.028.fcs</p></td><td colspan="1" rowspan="1"><p>C8</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>79.8</p></td><td colspan="1" rowspan="1"><p>3804</p></td><td colspan="1" rowspan="1"><p>1555</p></td><td colspan="1" rowspan="1"><p>103</p></td><td colspan="1" rowspan="1"><p>3634</p></td><td colspan="1" rowspan="1"><p>1508</p></td></tr><tr><td colspan="1" rowspan="1"><p>29: if142014-09-08.029.fcs</p></td><td colspan="1" rowspan="1"><p>C5</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>68.9</p></td><td colspan="1" rowspan="1"><p>3268</p></td><td colspan="1" rowspan="1"><p>1505</p></td><td colspan="1" rowspan="1"><p>95.7</p></td><td colspan="1" rowspan="1"><p>3128</p></td><td colspan="1" rowspan="1"><p>1419</p></td></tr><tr><td colspan="1" rowspan="1"><p>30: if142014-09-08.030.fcs</p></td><td colspan="1" rowspan="1"><p>C6</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>80.2</p></td><td colspan="1" rowspan="1"><p>3806</p></td><td colspan="1" rowspan="1"><p>1639</p></td><td colspan="1" rowspan="1"><p>110</p></td><td colspan="1" rowspan="1"><p>3630</p></td><td colspan="1" rowspan="1"><p>1541</p></td></tr><tr><td colspan="1" rowspan="1"><p>31: if142014-09-08.031.fcs</p></td><td colspan="1" rowspan="1"><p>C7</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>67.8</p></td><td colspan="1" rowspan="1"><p>3240</p></td><td colspan="1" rowspan="1"><p>1498</p></td><td colspan="1" rowspan="1"><p>91.7</p></td><td colspan="1" rowspan="1"><p>3041</p></td><td colspan="1" rowspan="1"><p>1412</p></td></tr><tr><td colspan="1" rowspan="1"><p>32: if142014-09-08.032.fcs</p></td><td colspan="1" rowspan="1"><p>C8</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>83.7</p></td><td colspan="1" rowspan="1"><p>3810</p></td><td colspan="1" rowspan="1"><p>1675</p></td><td colspan="1" rowspan="1"><p>107</p></td><td colspan="1" rowspan="1"><p>3618</p></td><td colspan="1" rowspan="1"><p>1621</p></td></tr><tr><td colspan="1" rowspan="1"><p>33: if142014-09-08.033.fcs</p></td><td colspan="1" rowspan="1"><p>AC1</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>0.895</p></td><td colspan="1" rowspan="1"><p>40.2</p></td><td colspan="1" rowspan="1"><p>16.8</p></td><td colspan="1" rowspan="1"><p>0.854</p></td><td colspan="1" rowspan="1"><p>34.9</p></td><td colspan="1" rowspan="1"><p>12.9</p></td></tr><tr><td colspan="1" rowspan="1"><p>34: if142014-09-08.034.fcs</p></td><td colspan="1" rowspan="1"><p>AC3</p></td><td colspan="1" rowspan="1"><p>+</p></td><td colspan="1" rowspan="1"><p>0.539</p></td><td colspan="1" rowspan="1"><p>28.4</p></td><td colspan="1" rowspan="1"><p>11</p></td><td colspan="1" rowspan="1"><p>0.785</p></td><td colspan="1" rowspan="1"><p>27.4</p></td><td colspan="1" rowspan="1"><p>10.7</p></td></tr><tr><td colspan="1" rowspan="1"><p>35: if142014-09-08.035.fcs</p></td><td colspan="1" rowspan="1"><p>AC1</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>0.464</p></td><td colspan="1" rowspan="1"><p>25.8</p></td><td colspan="1" rowspan="1"><p>9.39</p></td><td colspan="1" rowspan="1"><p>0.605</p></td><td colspan="1" rowspan="1"><p>25.9</p></td><td colspan="1" rowspan="1"><p>8.65</p></td></tr><tr><td colspan="1" rowspan="1"><p>36: if142014-09-08.036.fcs</p></td><td colspan="1" rowspan="1"><p>AC3</p></td><td colspan="1" rowspan="1"><p>-</p></td><td colspan="1" rowspan="1"><p>0.532</p></td><td colspan="1" rowspan="1"><p>28.3</p></td><td colspan="1" rowspan="1"><p>10.4</p></td><td colspan="1" rowspan="1"><p>0.661</p></td><td colspan="1" rowspan="1"><p>26.2</p></td><td colspan="1" rowspan="1"><p>9.23</p></td></tr></tbody></table><p></p><ul><li>Got ratios of fluorescence in +/- aTc and am reporting them in the table below</li></ul><a href="#" name="803fb1152dc946dc3b98f720d1637f36481109e8"></a><a href="#" name="5"></a><table cellpadding="0" cellspacing="0"><tbody><tr><td colspan="1" rowspan="1"><p>n/a</p></td><td colspan="1" rowspan="1"><p>n/a</p></td><td colspan="1" rowspan="1"><p>+/-</p></td><td colspan="1" rowspan="1"><p>smalls</p></td><td colspan="1" rowspan="1"><p></p></td><td colspan="1" rowspan="1"><p></p></td><td colspan="1" rowspan="1"><p>bugs</p></td><td colspan="1" rowspan="1"><p></p></td></tr><tr><td colspan="1" rowspan="1"><p>n/a</p></td><td colspan="1" rowspan="1"><p>n/a</p></td><td colspan="1" rowspan="1"><p>gfpa</p></td><td colspan="1" rowspan="1"><p>gfpa/fsca*1000</p></td><td colspan="1" rowspan="1"><p>gfpa/ssca*1000</p></td><td colspan="1" rowspan="1"><p>gfpa</p></td><td colspan="1" rowspan="1"><p>gfpa/fsca*1000</p></td><td colspan="1" rowspan="1"><p>gfpa/ssca*1000</p></td></tr><tr><td colspan="1" rowspan="1"><p>1: if142014-09-08.001.fcs</p></td><td colspan="1" rowspan="1"><p>G1</p></td><td colspan="1" rowspan="1"><p>0.771</p></td><td colspan="1" rowspan="1"><p>0.781</p></td><td colspan="1" rowspan="1"><p>0.772</p></td><td colspan="1" rowspan="1"><p>0.910</p></td><td colspan="1" rowspan="1"><p>0.797</p></td><td colspan="1" rowspan="1"><p>0.797</p></td></tr><tr><td colspan="1" rowspan="1"><p>2: if142014-09-08.002.fcs</p></td><td colspan="1" rowspan="1"><p>G2</p></td><td colspan="1" rowspan="1"><p>0.692</p></td><td colspan="1" rowspan="1"><p>0.671</p></td><td colspan="1" rowspan="1"><p>0.510</p></td><td colspan="1" rowspan="1"><p>0.982</p></td><td colspan="1" rowspan="1"><p>0.706</p></td><td colspan="1" rowspan="1"><p>0.526</p></td></tr><tr><td colspan="1" rowspan="1"><p>3: if142014-09-08.003.fcs</p></td><td colspan="1" rowspan="1"><p>G3</p></td><td colspan="1" rowspan="1"><p>0.717</p></td><td colspan="1" rowspan="1"><p>0.726</p></td><td colspan="1" rowspan="1"><p>0.593</p></td><td colspan="1" rowspan="1"><p>0.947</p></td><td colspan="1" rowspan="1"><p>0.757</p></td><td colspan="1" rowspan="1"><p>0.613</p></td></tr><tr><td colspan="1" rowspan="1"><p>4: if142014-09-08.004.fcs</p></td><td colspan="1" rowspan="1"><p>G4</p></td><td colspan="1" rowspan="1"><p>0.629</p></td><td colspan="1" rowspan="1"><p>0.641</p></td><td colspan="1" rowspan="1"><p>0.501</p></td><td colspan="1" rowspan="1"><p>0.782</p></td><td colspan="1" rowspan="1"><p>0.667</p></td><td colspan="1" rowspan="1"><p>0.518</p></td></tr><tr><td colspan="1" rowspan="1"><p>9: if142014-09-08.009.fcs</p></td><td colspan="1" rowspan="1"><p>C1</p></td><td colspan="1" rowspan="1"><p>1.144</p></td><td colspan="1" rowspan="1"><p>1.104</p></td><td colspan="1" rowspan="1"><p>1.208</p></td><td colspan="1" rowspan="1"><p>1.170</p></td><td colspan="1" rowspan="1"><p>1.122</p></td><td colspan="1" rowspan="1"><p>1.209</p></td></tr><tr><td colspan="1" rowspan="1"><p>10: if142014-09-08.010.fcs</p></td><td colspan="1" rowspan="1"><p>C2</p></td><td colspan="1" rowspan="1"><p>1.295</p></td><td colspan="1" rowspan="1"><p>1.317</p></td><td colspan="1" rowspan="1"><p>1.071</p></td><td colspan="1" rowspan="1"><p>1.359</p></td><td colspan="1" rowspan="1"><p>1.340</p></td><td colspan="1" rowspan="1"><p>1.036</p></td></tr><tr><td colspan="1" rowspan="1"><p>11: if142014-09-08.011.fcs</p></td><td colspan="1" rowspan="1"><p>C3</p></td><td colspan="1" rowspan="1"><p>1.010</p></td><td colspan="1" rowspan="1"><p>0.996</p></td><td colspan="1" rowspan="1"><p>1.006</p></td><td colspan="1" rowspan="1"><p>1.085</p></td><td colspan="1" rowspan="1"><p>1.018</p></td><td colspan="1" rowspan="1"><p>1.003</p></td></tr><tr><td colspan="1" rowspan="1"><p>12: if142014-09-08.012.fcs</p></td><td colspan="1" rowspan="1"><p>C4</p></td><td colspan="1" rowspan="1"><p>1.311</p></td><td colspan="1" rowspan="1"><p>1.350</p></td><td colspan="1" rowspan="1"><p>1.214</p></td><td colspan="1" rowspan="1"><p>1.201</p></td><td colspan="1" rowspan="1"><p>1.424</p></td><td colspan="1" rowspan="1"><p>1.232</p></td></tr><tr><td colspan="1" rowspan="1"><p>17: if142014-09-08.017.fcs</p></td><td colspan="1" rowspan="1"><p>G5</p></td><td colspan="1" rowspan="1"><p>0.421</p></td><td colspan="1" rowspan="1"><p>0.440</p></td><td colspan="1" rowspan="1"><p>0.394</p></td><td colspan="1" rowspan="1"><p>0.432</p></td><td colspan="1" rowspan="1"><p>0.459</p></td><td colspan="1" rowspan="1"><p>0.401</p></td></tr><tr><td colspan="1" rowspan="1"><p>18: if142014-09-08.018.fcs</p></td><td colspan="1" rowspan="1"><p>G6</p></td><td colspan="1" rowspan="1"><p>0.506</p></td><td colspan="1" rowspan="1"><p>0.500</p></td><td colspan="1" rowspan="1"><p>0.498</p></td><td colspan="1" rowspan="1"><p>0.548</p></td><td colspan="1" rowspan="1"><p>0.503</p></td><td colspan="1" rowspan="1"><p>0.498</p></td></tr><tr><td colspan="1" rowspan="1"><p>19: if142014-09-08.019.fcs</p></td><td colspan="1" rowspan="1"><p>G7</p></td><td colspan="1" rowspan="1"><p>0.438</p></td><td colspan="1" rowspan="1"><p>0.456</p></td><td colspan="1" rowspan="1"><p>0.400</p></td><td colspan="1" rowspan="1"><p>0.449</p></td><td colspan="1" rowspan="1"><p>0.470</p></td><td colspan="1" rowspan="1"><p>0.413</p></td></tr><tr><td colspan="1" rowspan="1"><p>20: if142014-09-08.020.fcs</p></td><td colspan="1" rowspan="1"><p>G8</p></td><td colspan="1" rowspan="1"><p>0.415</p></td><td colspan="1" rowspan="1"><p>0.431</p></td><td colspan="1" rowspan="1"><p>0.376</p></td><td colspan="1" rowspan="1"><p>0.427</p></td><td colspan="1" rowspan="1"><p>0.450</p></td><td colspan="1" rowspan="1"><p>0.390</p></td></tr><tr><td colspan="1" rowspan="1"><p>25: if142014-09-08.025.fcs</p></td><td colspan="1" rowspan="1"><p>C5</p></td><td colspan="1" rowspan="1"><p>1.155</p></td><td colspan="1" rowspan="1"><p>1.211</p></td><td colspan="1" rowspan="1"><p>0.987</p></td><td colspan="1" rowspan="1"><p>1.181</p></td><td colspan="1" rowspan="1"><p>1.219</p></td><td colspan="1" rowspan="1"><p>0.968</p></td></tr><tr><td colspan="1" rowspan="1"><p>26: if142014-09-08.026.fcs</p></td><td colspan="1" rowspan="1"><p>C6</p></td><td colspan="1" rowspan="1"><p>1.007</p></td><td colspan="1" rowspan="1"><p>1.063</p></td><td colspan="1" rowspan="1"><p>0.924</p></td><td colspan="1" rowspan="1"><p>1.027</p></td><td colspan="1" rowspan="1"><p>1.114</p></td><td colspan="1" rowspan="1"><p>0.925</p></td></tr><tr><td colspan="1" rowspan="1"><p>27: if142014-09-08.027.fcs</p></td><td colspan="1" rowspan="1"><p>C7</p></td><td colspan="1" rowspan="1"><p>1.190</p></td><td colspan="1" rowspan="1"><p>1.251</p></td><td colspan="1" rowspan="1"><p>0.995</p></td><td colspan="1" rowspan="1"><p>1.265</p></td><td colspan="1" rowspan="1"><p>1.289</p></td><td colspan="1" rowspan="1"><p>0.974</p></td></tr><tr><td colspan="1" rowspan="1"><p>28: if142014-09-08.028.fcs</p></td><td colspan="1" rowspan="1"><p>C8</p></td><td colspan="1" rowspan="1"><p>0.953</p></td><td colspan="1" rowspan="1"><p>0.998</p></td><td colspan="1" rowspan="1"><p>0.928</p></td><td colspan="1" rowspan="1"><p>0.963</p></td><td colspan="1" rowspan="1"><p>1.004</p></td><td colspan="1" rowspan="1"><p>0.930</p></td></tr><tr><td colspan="1" rowspan="1"><p>33: if142014-09-08.033.fcs</p></td><td colspan="1" rowspan="1"><p>AC1</p></td><td colspan="1" rowspan="1"><p>1.929</p></td><td colspan="1" rowspan="1"><p>1.558</p></td><td colspan="1" rowspan="1"><p>1.789</p></td><td colspan="1" rowspan="1"><p>1.412</p></td><td colspan="1" rowspan="1"><p>1.347</p></td><td colspan="1" rowspan="1"><p>1.491</p></td></tr><tr><td colspan="1" rowspan="1"><p>34: if142014-09-08.034.fcs</p></td><td colspan="1" rowspan="1"><p>AC3</p></td><td colspan="1" rowspan="1"><p>1.013</p></td><td colspan="1" rowspan="1"><p>1.004</p></td><td colspan="1" rowspan="1"><p>1.058</p></td><td colspan="1" rowspan="1"><p>1.188</p></td><td colspan="1" rowspan="1"><p>1.046</p></td><td colspan="1" rowspan="1"><p>1.159</p></td></tr></tbody></table><ul><li></li></ul><hr style="page-break-before:always;display:none;"><p><span style="overflow: hidden; display: inline-block; margin: 0.00px 0.00px; border: 0.00px solid #000000; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px); width: 560.00px; height: 420.00px;"><img alt="" src="images/image19.jpg" style="width: 560.00px; height: 420.00px; margin-left: 0.00px; margin-top: 0.00px; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px);" title=""></p><p>Box plot of just G1-4 and C1-4. Note that the G clones from plates (clones 5-8) have even smaller ratios. </p><hr style="page-break-before:always;display:none;"><p></p><p></p><p></p><p></p><hr style="page-break-before:always;display:none;"><p></p><p></p><p></p><p></p><h2><a name="h.h7pf932a50dt"></a>9/9/14</h2><p>Objective: assembly decoy binding site arrays</p><ul><li>Garima sets up a PCR and is asked to fill in notes about it.</li><li>Results</li><li>Charlie sets up a new set of tubes for PCR with a 5 rx master mix containing</li></ul><ul><li>190uL H2O</li><li>50 uL Q5 buffer</li><li>2.0 uL oligo 1 (gfp1-spacer-gfp1)</li><li>2.5 uL Q5 poly</li><li>5 uL dNTPs</li></ul><ul><li>And then divides that in to four tubes before adding in the following volumes of oligo 2 (spacer-gfp1-space)</li></ul><a href="#" name="fd71260aa00de3f043dfc426450b7d96e13e45b0"></a><a href="#" name="6"></a><table cellpadding="0" cellspacing="0"><tbody><tr><td colspan="1" rowspan="1"><p>tube number</p></td><td colspan="1" rowspan="1"><p>1</p></td><td colspan="1" rowspan="1"><p>2</p></td><td colspan="1" rowspan="1"><p>3</p></td><td colspan="1" rowspan="1"><p>4</p></td></tr><tr><td colspan="1" rowspan="1"><p>uL oligo 2</p></td><td colspan="1" rowspan="1"><p>0</p></td><td colspan="1" rowspan="1"><p>0.5</p></td><td colspan="1" rowspan="1"><p>1.0</p></td><td colspan="1" rowspan="1"><p>2.0</p></td></tr></tbody></table><ul><li><span style="overflow: hidden; display: inline-block; margin: 0.00px 0.00px; border: 0.00px solid #000000; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px); width: 624.00px; height: 468.00px;"><img alt="Sep 10, 2014 1:24:31 PM.jpg" src="images/image04.jpg" style="width: 624.00px; height: 468.00px; margin-left: 0.00px; margin-top: 0.00px; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px);" title=""></li></ul><ul><li>Streakiness on the row with 0.5uL of oligo 2 looks promising? IGEM protocol on the thermal cycler modified</li></ul><h2><a name="h.3838tuwxw3ds"></a>
| + | <ul><li>Started colonies from plates and diluted in to +/- aTc this morning. Let shake for 5(?) hours before collecting 500uL and spin/resusp in PBS for flow.</li><li>Took data to flowjo and plotted histograms below. This is ungated fluorescence</li><li></li><li><span style="overflow: hidden; display: inline-block; margin: 0.00px 0.00px; border: 0.00px solid #000000; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px); width: 624.00px; height: 482.67px;"><img alt="" src="images/image44.jpg" style="width: 624.00px; height: 482.67px; margin-left: 0.00px; margin-top: 0.00px; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px);" title=""></li><li>table with mean GFPAs</li></ul><a href="#" name="096a26566c8f532b5e11f6f110d57e6edec006da"></a><a href="#" name="3"></a><table cellpadding="0" cellspacing="0"><tbody><tr><td><p></p></td><td><p></p></td><td><p></p></td><td><p>ungated GFPA</p></td><td><p>+/- aTc</p></td></tr><tr><td><p>1: if142014-09-04.001.fcs</p></td><td><p>AC-1</p></td><td><p>+ aTc</p></td><td><p>0.402</p></td><td><p>1.185840708</p></td></tr><tr><td><p>2: if142014-09-04.002.fcs</p></td><td><p>AC-2</p></td><td><p>+ aTc</p></td><td><p>0.455</p></td><td><p>1.197368421</p></td></tr><tr><td><p>3: if142014-09-04.003.fcs</p></td><td><p>G-1</p></td><td><p>+ aTc</p></td><td><p>14.9</p></td><td><p>0.949044586</p></td></tr><tr><td><p>4: if142014-09-04.004.fcs</p></td><td><p>G-2</p></td><td><p>+ aTc</p></td><td><p>11.7</p></td><td><p>0.573529412</p></td></tr><tr><td><p>5: if142014-09-04.005.fcs</p></td><td><p>G-3</p></td><td><p>+ aTc</p></td><td><p>8.91</p></td><td><p>0.530357143</p></td></tr><tr><td><p>6: if142014-09-04.006.fcs</p></td><td><p>G-4</p></td><td><p>+ aTc</p></td><td><p>5.92</p></td><td><p>0.34619883</p></td></tr><tr><td><p>7: if142014-09-04.007.fcs</p></td><td><p>C-1</p></td><td><p>+ aTc</p></td><td><p>186</p></td><td><p>1.016393443</p></td></tr><tr><td><p>8: if142014-09-04.008.fcs</p></td><td><p>C-2</p></td><td><p>+ aTc</p></td><td><p>224</p></td><td><p>1.230769231</p></td></tr><tr><td><p>9: if142014-09-04.009.fcs</p></td><td><p>C-3</p></td><td><p>+ aTc</p></td><td><p>214</p></td><td><p>0.922413793</p></td></tr><tr><td><p>10: if142014-09-04.010.fcs</p></td><td><p>C-4</p></td><td><p>+ aTc</p></td><td><p>222</p></td><td><p>1.193548387</p></td></tr><tr><td><p>11: if142014-09-04.011.fcs</p></td><td><p>AC-1</p></td><td><p>- aTc</p></td><td><p>0.339</p></td><td><p></p></td></tr><tr><td><p>12: if142014-09-04.012.fcs</p></td><td><p>AC-2</p></td><td><p>- aTc</p></td><td><p>0.38</p></td><td><p></p></td></tr><tr><td><p>13: if142014-09-04.013.fcs</p></td><td><p>G-1</p></td><td><p>- aTc</p></td><td><p>15.7</p></td><td><p></p></td></tr><tr><td><p>14: if142014-09-04.014.fcs</p></td><td><p>G-2</p></td><td><p>- aTc</p></td><td><p>20.4</p></td><td><p></p></td></tr><tr><td><p>15: if142014-09-04.015.fcs</p></td><td><p>G-3</p></td><td><p>- aTc</p></td><td><p>16.8</p></td><td><p></p></td></tr><tr><td><p>16: if142014-09-04.016.fcs</p></td><td><p>G-4</p></td><td><p>- aTc</p></td><td><p>17.1</p></td><td><p></p></td></tr><tr><td><p>17: if142014-09-04.017.fcs</p></td><td><p>C-1</p></td><td><p>- aTc</p></td><td><p>183</p></td><td><p></p></td></tr><tr><td><p>18: if142014-09-04.018.fcs</p></td><td><p>C-2</p></td><td><p>- aTc</p></td><td><p>182</p></td><td><p></p></td></tr><tr><td><p>19: if142014-09-04.019.fcs</p></td><td><p>C-3</p></td><td><p>- aTc</p></td><td><p>232</p></td><td><p></p></td></tr><tr><td><p>20: if142014-09-04.020.fcs</p></td><td><p>C-4</p></td><td><p>- aTc</p></td><td><p>186</p></td><td><p></p></td></tr></tbody></table><ul><li>aTc treatment seems to be moving fluor in the opposite direction of what we’d expect of antitracr was doing its job. The degree is inconsistent, reflow is called for. </li></ul><p></p><p></p><p></p><p></p><hr style="page-break-before:always;display:none;"><p></p><hr style="page-break-before:always;display:none;"><p></p><p></p><h2><a name="h.ohhk3mdbkmcm"></a>9/8/14</h2><p>Objective: retest backbone shuffled antitracr bacteria</p><ul><li>Yesterday picked 4 new colonies from plates and 4 from 9/4/14 frozen stocks and grew in LB + antibiotics overnight. Diluted in to +/- aTc this morning and let shake for 5(?) hours before collecting 500uL and spin/resusp in PBS</li><li>G1-4 are old frozen coli with K608012, pdCas9-GFP1, and 1A2-R0040-antitracr, C1-4 are old frozen with pdCas9. clones 5-8 are new from plates. AC are no-fluor (1A2 and pdCas9) but they were diluted in to triple antibiotic medium instead of double so they did not grow and are not reliable indicators of bg fluorescence on this run.</li><li>After flow, isolated singlets with flowjo (smalls) or left alone (bugs) and got gfpa unmodified or normalized by fsca or ssca. </li><li></li></ul><a href="#" name="777f62c089f7173b14ec6d88236db6c3e53afff8"></a><a href="#" name="4"></a><table cellpadding="0" cellspacing="0"<><tbody><tr><td><p></p></td><td><p></p></td><td><p></p></td><td><p></p></td><td><p>smalls</p></td><td><p></p></td><td><p></p></td><td><p>bugs</p></td><td><p></p></td></tr><tr><td><p></p></td><td><p></p></td><td><p>aTc</p></td><td><p>gfpa</p></td><td><p>gfpa/fsca*1000</p></td><td><p>gfpa/ssca*1000</p></td><td><p>gfpa</p></td><td><p>gfpa/fsca*1000</p></td><td><p>gfpa/ssca*1000</p></td></tr><tr><td><p>1: if142014-09-08.001.fcs</p></td><td><p>G1</p></td><td><p>+</p></td><td><p>5.25</p></td><td><p>243</p></td><td><p>112</p></td><td><p>7.79</p></td><td><p>247</p></td><td><p>114</p></td></tr><tr><td><p>2: if142014-09-08.002.fcs</p></td><td><p>G2</p></td><td><p>+</p></td><td><p>4.41</p></td><td><p>226</p></td><td><p>78.1</p></td><td><p>7.12</p></td><td><p>228</p></td><td><p>76.2</p></td></tr><tr><td><p>3: if142014-09-08.003.fcs</p></td><td><p>G3</p></td><td><p>+</p></td><td><p>4.66</p></td><td><p>231</p></td><td><p>84.8</p></td><td><p>7.8</p></td><td><p>231</p></td><td><p>83.4</p></td></tr><tr><td><p>4: if142014-09-08.004.fcs</p></td><td><p>G4</p></td><td><p>+</p></td><td><p>3.86</p></td><td><p>191</p></td><td><p>70.1</p></td><td><p>5.99</p></td><td><p>192</p></td><td><p>67.9</p></td></tr><tr><td><p>5: if142014-09-08.005.fcs</p></td><td><p>G1</p></td><td><p>-</p></td><td><p>6.81</p></td><td><p>311</p></td><td><p>145</p></td><td><p>8.56</p></td><td><p>310</p></td><td><p>143</p></td></tr><tr><td><p>6: if142014-09-08.006.fcs</p></td><td><p>G2</p></td><td><p>-</p></td><td><p>6.37</p></td><td><p>337</p></td><td><p>153</p></td><td><p>7.25</p></td><td><p>323</p></td><td><p>145</p></td></tr><tr><td><p>7: if142014-09-08.007.fcs</p></td><td><p>G3</p></td><td><p>-</p></td><td><p>6.5</p></td><td><p>318</p></td><td><p>143</p></td><td><p>8.24</p></td><td><p>305</p></td><td><p>136</p></td></tr><tr><td><p>8: if142014-09-08.008.fcs</p></td><td><p>G4</p></td><td><p>-</p></td><td><p>6.14</p></td><td><p>298</p></td><td><p>140</p></td><td><p>7.66</p></td><td><p>288</p></td><td><p>131</p></td></tr><tr><td><p>9: if142014-09-08.009.fcs</p></td><td><p>C1</p></td><td><p>+</p></td><td><p>88.9</p></td><td><p>4071</p></td><td><p>1777</p></td><td><p>117</p></td><td><p>3978</p></td><td><p>1706</p></td></tr><tr><td><p>10: if142014-09-08.010.fcs</p></td><td><p>C2</p></td><td><p>+</p></td><td><p>83.5</p></td><td><p>4073</p></td><td><p>1547</p></td><td><p>120</p></td><td><p>3969</p></td><td><p>1422</p></td></tr><tr><td><p>11: if142014-09-08.011.fcs</p></td><td><p>C3</p></td><td><p>+</p></td><td><p>81.7</p></td><td><p>3957</p></td><td><p>1551</p></td><td><p>115</p></td><td><p>3840</p></td><td><p>1437</p></td></tr><tr><td><p>12: if142014-09-08.012.fcs</p></td><td><p>C4</p></td><td><p>+</p></td><td><p>77.2</p></td><td><p>3915</p></td><td><p>1577</p></td><td><p>95.7</p></td><td><p>3883</p></td><td><p>1503</p></td></tr><tr><td><p>13: if142014-09-08.013.fcs</p></td><td><p>C1</p></td><td><p>-</p></td><td><p>77.7</p></td><td><p>3686</p></td><td><p>1471</p></td><td><p>100</p></td><td><p>3545</p></td><td><p>1411</p></td></tr><tr><td><p>14: if142014-09-08.014.fcs</p></td><td><p>C2</p></td><td><p>-</p></td><td><p>64.5</p></td><td><p>3093</p></td><td><p>1445</p></td><td><p>88.3</p></td><td><p>2962</p></td><td><p>1373</p></td></tr><tr><td><p>15: if142014-09-08.015.fcs</p></td><td><p>C3</p></td><td><p>-</p></td><td><p>80.9</p></td><td><p>3971</p></td><td><p>1541</p></td><td><p>106</p></td><td><p>3771</p></td><td><p>1432</p></td></tr><tr><td><p>16: if142014-09-08.016.fcs</p></td><td><p>C4</p></td><td><p>-</p></td><td><p>58.9</p></td><td><p>2899</p></td><td><p>1299</p></td><td><p>79.7</p></td><td><p>2727</p></td><td><p>1220</p></td></tr><tr><td><p>17: if142014-09-08.017.fcs</p></td><td><p>G5</p></td><td><p>+</p></td><td><p>4.1</p></td><td><p>210</p></td><td><p>79.1</p></td><td><p>5.4</p></td><td><p>209</p></td><td><p>76.2</p></td></tr><tr><td><p>18: if142014-09-08.018.fcs</p></td><td><p>G6</p></td><td><p>+</p></td><td><p>4.9</p></td><td><p>225</p></td><td><p>98.2</p></td><td><p>7.07</p></td><td><p>226</p></td><td><p>97.6</p></td></tr><tr><td><p>19: if142014-09-08.019.fcs</p></td><td><p>G7</p></td><td><p>+</p></td><td><p>4.42</p></td><td><p>220</p></td><td><p>84.7</p></td><td><p>5.97</p></td><td><p>218</p></td><td><p>81.7</p></td></tr><tr><td><p>20: if142014-09-08.020.fcs</p></td><td><p>G8</p></td><td><p>+</p></td><td><p>4.19</p></td><td><p>211</p></td><td><p>81.3</p></td><td><p>5.68</p></td><td><p>213</p></td><td><p>79.5</p></td></tr><tr><td><p>21: if142014-09-08.021.fcs</p></td><td><p>G5</p></td><td><p>-</p></td><td><p>9.73</p></td><td><p>477</p></td><td><p>201</p></td><td><p>12.5</p></td><td><p>455</p></td><td><p>190</p></td></tr><tr><td><p>22: if142014-09-08.022.fcs</p></td><td><p>G6</p></td><td><p>-</p></td><td><p>9.68</p></td><td><p>450</p></td><td><p>197</p></td><td><p>12.9</p></td><td><p>449</p></td><td><p>196</p></td></tr><tr><td><p>23: if142014-09-08.023.fcs</p></td><td><p>G7</p></td><td><p>-</p></td><td><p>10.1</p></td><td><p>482</p></td><td><p>212</p></td><td><p>13.3</p></td><td><p>464</p></td><td><p>198</p></td></tr><tr><td><p>24: if142014-09-08.024.fcs</p></td><td><p>G8</p></td><td><p>-</p></td><td><p>10.1</p></td><td><p>489</p></td><td><p>216</p></td><td><p>13.3</p></td><td><p>473</p></td><td><p>204</p></td></tr><tr><td><p>25: if142014-09-08.025.fcs</p></td><td><p>C5</p></td><td><p>+</p></td><td><p>79.6</p></td><td><p>3958</p></td><td><p>1486</p></td><td><p>113</p></td><td><p>3813</p></td><td><p>1374</p></td></tr><tr><td><p>26: if142014-09-08.026.fcs</p></td><td><p>C6</p></td><td><p>+</p></td><td><p>80.8</p></td><td><p>4045</p></td><td><p>1514</p></td><td><p>113</p></td><td><p>4043</p></td><td><p>1425</p></td></tr><tr><td><p>27: if142014-09-08.027.fcs</p></td><td><p>C7</p></td><td><p>+</p></td><td><p>80.7</p></td><td><p>4053</p></td><td><p>1491</p></td><td><p>116</p></td><td><p>3921</p></td><td><p>1375</p></td></tr><tr><td><p>28: if142014-09-08.028.fcs</p></td><td><p>C8</p></td><td><p>+</p></td><td><p>79.8</p></td><td><p>3804</p></td><td><p>1555</p></td><td><p>103</p></td><td><p>3634</p></td><td><p>1508</p></td></tr><tr><td><p>29: if142014-09-08.029.fcs</p></td><td><p>C5</p></td><td><p>-</p></td><td><p>68.9</p></td><td><p>3268</p></td><td><p>1505</p></td><td><p>95.7</p></td><td><p>3128</p></td><td><p>1419</p></td></tr><tr><td><p>30: if142014-09-08.030.fcs</p></td><td><p>C6</p></td><td><p>-</p></td><td><p>80.2</p></td><td><p>3806</p></td><td><p>1639</p></td><td><p>110</p></td><td><p>3630</p></td><td><p>1541</p></td></tr><tr><td><p>31: if142014-09-08.031.fcs</p></td><td><p>C7</p></td><td><p>-</p></td><td><p>67.8</p></td><td><p>3240</p></td><td><p>1498</p></td><td><p>91.7</p></td><td><p>3041</p></td><td><p>1412</p></td></tr><tr><td><p>32: if142014-09-08.032.fcs</p></td><td><p>C8</p></td><td><p>-</p></td><td><p>83.7</p></td><td><p>3810</p></td><td><p>1675</p></td><td><p>107</p></td><td><p>3618</p></td><td><p>1621</p></td></tr><tr><td><p>33: if142014-09-08.033.fcs</p></td><td><p>AC1</p></td><td><p>+</p></td><td><p>0.895</p></td><td><p>40.2</p></td><td><p>16.8</p></td><td><p>0.854</p></td><td><p>34.9</p></td><td><p>12.9</p></td></tr><tr><td><p>34: if142014-09-08.034.fcs</p></td><td><p>AC3</p></td><td><p>+</p></td><td><p>0.539</p></td><td><p>28.4</p></td><td><p>11</p></td><td><p>0.785</p></td><td><p>27.4</p></td><td><p>10.7</p></td></tr><tr><td><p>35: if142014-09-08.035.fcs</p></td><td><p>AC1</p></td><td><p>-</p></td><td><p>0.464</p></td><td><p>25.8</p></td><td><p>9.39</p></td><td><p>0.605</p></td><td><p>25.9</p></td><td><p>8.65</p></td></tr><tr><td><p>36: if142014-09-08.036.fcs</p></td><td><p>AC3</p></td><td><p>-</p></td><td><p>0.532</p></td><td><p>28.3</p></td><td><p>10.4</p></td><td><p>0.661</p></td><td><p>26.2</p></td><td><p>9.23</p></td></tr></tbody></table><p></p><ul><li>Got ratios of fluorescence in +/- aTc and am reporting them in the table below</li></ul><a href="#" name="803fb1152dc946dc3b98f720d1637f36481109e8"></a><a href="#" name="5"></a><table cellpadding="0" cellspacing="0"><tbody><tr><td><p>n/a</p></td><td><p>n/a</p></td><td><p>+/-</p></td><td><p>smalls</p></td><td><p></p></td><td><p></p></td><td><p>bugs</p></td><td><p></p></td></tr><tr><td><p>n/a</p></td><td><p>n/a</p></td><td><p>gfpa</p></td><td><p>gfpa/fsca*1000</p></td><td><p>gfpa/ssca*1000</p></td><td><p>gfpa</p></td><td><p>gfpa/fsca*1000</p></td><td><p>gfpa/ssca*1000</p></td></tr><tr><td><p>1: if142014-09-08.001.fcs</p></td><td><p>G1</p></td><td><p>0.771</p></td><td><p>0.781</p></td><td><p>0.772</p></td><td><p>0.910</p></td><td><p>0.797</p></td><td><p>0.797</p></td></tr><tr><td><p>2: if142014-09-08.002.fcs</p></td><td><p>G2</p></td><td><p>0.692</p></td><td><p>0.671</p></td><td><p>0.510</p></td><td><p>0.982</p></td><td><p>0.706</p></td><td><p>0.526</p></td></tr><tr><td><p>3: if142014-09-08.003.fcs</p></td><td><p>G3</p></td><td><p>0.717</p></td><td><p>0.726</p></td><td><p>0.593</p></td><td><p>0.947</p></td><td><p>0.757</p></td><td><p>0.613</p></td></tr><tr><td><p>4: if142014-09-08.004.fcs</p></td><td><p>G4</p></td><td><p>0.629</p></td><td><p>0.641</p></td><td><p>0.501</p></td><td><p>0.782</p></td><td><p>0.667</p></td><td><p>0.518</p></td></tr><tr><td><p>9: if142014-09-08.009.fcs</p></td><td><p>C1</p></td><td><p>1.144</p></td><td><p>1.104</p></td><td><p>1.208</p></td><td><p>1.170</p></td><td><p>1.122</p></td><td><p>1.209</p></td></tr><tr><td><p>10: if142014-09-08.010.fcs</p></td><td><p>C2</p></td><td><p>1.295</p></td><td><p>1.317</p></td><td><p>1.071</p></td><td><p>1.359</p></td><td><p>1.340</p></td><td><p>1.036</p></td></tr><tr><td><p>11: if142014-09-08.011.fcs</p></td><td><p>C3</p></td><td><p>1.010</p></td><td><p>0.996</p></td><td><p>1.006</p></td><td><p>1.085</p></td><td><p>1.018</p></td><td><p>1.003</p></td></tr><tr><td><p>12: if142014-09-08.012.fcs</p></td><td><p>C4</p></td><td><p>1.311</p></td><td><p>1.350</p></td><td><p>1.214</p></td><td><p>1.201</p></td><td><p>1.424</p></td><td><p>1.232</p></td></tr><tr><td><p>17: if142014-09-08.017.fcs</p></td><td><p>G5</p></td><td><p>0.421</p></td><td><p>0.440</p></td><td><p>0.394</p></td><td><p>0.432</p></td><td><p>0.459</p></td><td><p>0.401</p></td></tr><tr><td><p>18: if142014-09-08.018.fcs</p></td><td><p>G6</p></td><td><p>0.506</p></td><td><p>0.500</p></td><td><p>0.498</p></td><td><p>0.548</p></td><td><p>0.503</p></td><td><p>0.498</p></td></tr><tr><td><p>19: if142014-09-08.019.fcs</p></td><td><p>G7</p></td><td><p>0.438</p></td><td><p>0.456</p></td><td><p>0.400</p></td><td><p>0.449</p></td><td><p>0.470</p></td><td><p>0.413</p></td></tr><tr><td><p>20: if142014-09-08.020.fcs</p></td><td><p>G8</p></td><td><p>0.415</p></td><td><p>0.431</p></td><td><p>0.376</p></td><td><p>0.427</p></td><td><p>0.450</p></td><td><p>0.390</p></td></tr><tr><td><p>25: if142014-09-08.025.fcs</p></td><td><p>C5</p></td><td><p>1.155</p></td><td><p>1.211</p></td><td><p>0.987</p></td><td><p>1.181</p></td><td><p>1.219</p></td><td><p>0.968</p></td></tr><tr><td><p>26: if142014-09-08.026.fcs</p></td><td><p>C6</p></td><td><p>1.007</p></td><td><p>1.063</p></td><td><p>0.924</p></td><td><p>1.027</p></td><td><p>1.114</p></td><td><p>0.925</p></td></tr><tr><td><p>27: if142014-09-08.027.fcs</p></td><td><p>C7</p></td><td><p>1.190</p></td><td><p>1.251</p></td><td><p>0.995</p></td><td><p>1.265</p></td><td><p>1.289</p></td><td><p>0.974</p></td></tr><tr><td><p>28: if142014-09-08.028.fcs</p></td><td><p>C8</p></td><td><p>0.953</p></td><td><p>0.998</p></td><td><p>0.928</p></td><td><p>0.963</p></td><td><p>1.004</p></td><td><p>0.930</p></td></tr><tr><td><p>33: if142014-09-08.033.fcs</p></td><td><p>AC1</p></td><td><p>1.929</p></td><td><p>1.558</p></td><td><p>1.789</p></td><td><p>1.412</p></td><td><p>1.347</p></td><td><p>1.491</p></td></tr><tr><td><p>34: if142014-09-08.034.fcs</p></td><td><p>AC3</p></td><td><p>1.013</p></td><td><p>1.004</p></td><td><p>1.058</p></td><td><p>1.188</p></td><td><p>1.046</p></td><td><p>1.159</p></td></tr></tbody></table><ul><li></li></ul><hr style="page-break-before:always;display:none;"><p><span style="overflow: hidden; display: inline-block; margin: 0.00px 0.00px; border: 0.00px solid #000000; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px); width: 560.00px; height: 420.00px;"><img alt="" src="images/image19.jpg" style="width: 560.00px; height: 420.00px; margin-left: 0.00px; margin-top: 0.00px; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px);" title=""></p><p>Box plot of just G1-4 and C1-4. Note that the G clones from plates (clones 5-8) have even smaller ratios. </p><hr style="page-break-before:always;display:none;"><p></p><p></p><p></p><p></p><hr style="page-break-before:always;display:none;"><p></p><p></p><p></p><p></p><h2><a name="h.h7pf932a50dt"></a>9/9/14</h2><p>Objective: assembly decoy binding site arrays</p><ul><li>Garima sets up a PCR and is asked to fill in notes about it.</li><li>Results</li><li>Charlie sets up a new set of tubes for PCR with a 5 rx master mix containing</li></ul><ul><li>190uL H2O</li><li>50 uL Q5 buffer</li><li>2.0 uL oligo 1 (gfp1-spacer-gfp1)</li><li>2.5 uL Q5 poly</li><li>5 uL dNTPs</li></ul><ul><li>And then divides that in to four tubes before adding in the following volumes of oligo 2 (spacer-gfp1-space)</li></ul><a href="#" name="fd71260aa00de3f043dfc426450b7d96e13e45b0"></a><a href="#" name="6"></a><table cellpadding="0" cellspacing="0"><tbody><tr><td><p>tube number</p></td><td><p>1</p></td><td><p>2</p></td><td><p>3</p></td><td><p>4</p></td></tr><tr><td><p>uL oligo 2</p></td><td><p>0</p></td><td><p>0.5</p></td><td><p>1.0</p></td><td><p>2.0</p></td></tr></tbody></table><ul><li><span style="overflow: hidden; display: inline-block; margin: 0.00px 0.00px; border: 0.00px solid #000000; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px); width: 624.00px; height: 468.00px;"><img alt="Sep 10, 2014 1:24:31 PM.jpg" src="images/image04.jpg" style="width: 624.00px; height: 468.00px; margin-left: 0.00px; margin-top: 0.00px; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px);" title=""></li></ul><ul><li>Streakiness on the row with 0.5uL of oligo 2 looks promising? IGEM protocol on the thermal cycler modified</li></ul><h2><a name="h.3838tuwxw3ds"></a> |
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Revision as of 04:12, 17 October 2014
9/4/14
Objective: test backbone shuffled antitracr bacteria
- Started colonies from plates and diluted in to +/- aTc this morning. Let shake for 5(?) hours before collecting 500uL and spin/resusp in PBS for flow.
- Took data to flowjo and plotted histograms below. This is ungated fluorescence
- table with mean GFPAs
| | | ungated GFPA | +/- aTc |
1: if142014-09-04.001.fcs | AC-1 | + aTc | 0.402 | 1.185840708 |
2: if142014-09-04.002.fcs | AC-2 | + aTc | 0.455 | 1.197368421 |
3: if142014-09-04.003.fcs | G-1 | + aTc | 14.9 | 0.949044586 |
4: if142014-09-04.004.fcs | G-2 | + aTc | 11.7 | 0.573529412 |
5: if142014-09-04.005.fcs | G-3 | + aTc | 8.91 | 0.530357143 |
6: if142014-09-04.006.fcs | G-4 | + aTc | 5.92 | 0.34619883 |
7: if142014-09-04.007.fcs | C-1 | + aTc | 186 | 1.016393443 |
8: if142014-09-04.008.fcs | C-2 | + aTc | 224 | 1.230769231 |
9: if142014-09-04.009.fcs | C-3 | + aTc | 214 | 0.922413793 |
10: if142014-09-04.010.fcs | C-4 | + aTc | 222 | 1.193548387 |
11: if142014-09-04.011.fcs | AC-1 | - aTc | 0.339 | |
12: if142014-09-04.012.fcs | AC-2 | - aTc | 0.38 | |
13: if142014-09-04.013.fcs | G-1 | - aTc | 15.7 | |
14: if142014-09-04.014.fcs | G-2 | - aTc | 20.4 | |
15: if142014-09-04.015.fcs | G-3 | - aTc | 16.8 | |
16: if142014-09-04.016.fcs | G-4 | - aTc | 17.1 | |
17: if142014-09-04.017.fcs | C-1 | - aTc | 183 | |
18: if142014-09-04.018.fcs | C-2 | - aTc | 182 | |
19: if142014-09-04.019.fcs | C-3 | - aTc | 232 | |
20: if142014-09-04.020.fcs | C-4 | - aTc | 186 | |
- aTc treatment seems to be moving fluor in the opposite direction of what we’d expect of antitracr was doing its job. The degree is inconsistent, reflow is called for.
9/8/14
Objective: retest backbone shuffled antitracr bacteria
- Yesterday picked 4 new colonies from plates and 4 from 9/4/14 frozen stocks and grew in LB + antibiotics overnight. Diluted in to +/- aTc this morning and let shake for 5(?) hours before collecting 500uL and spin/resusp in PBS
- G1-4 are old frozen coli with K608012, pdCas9-GFP1, and 1A2-R0040-antitracr, C1-4 are old frozen with pdCas9. clones 5-8 are new from plates. AC are no-fluor (1A2 and pdCas9) but they were diluted in to triple antibiotic medium instead of double so they did not grow and are not reliable indicators of bg fluorescence on this run.
- After flow, isolated singlets with flowjo (smalls) or left alone (bugs) and got gfpa unmodified or normalized by fsca or ssca.
| | | | smalls | | | bugs | |
| | aTc | gfpa | gfpa/fsca*1000 | gfpa/ssca*1000 | gfpa | gfpa/fsca*1000 | gfpa/ssca*1000 |
1: if142014-09-08.001.fcs | G1 | + | 5.25 | 243 | 112 | 7.79 | 247 | 114 |
2: if142014-09-08.002.fcs | G2 | + | 4.41 | 226 | 78.1 | 7.12 | 228 | 76.2 |
3: if142014-09-08.003.fcs | G3 | + | 4.66 | 231 | 84.8 | 7.8 | 231 | 83.4 |
4: if142014-09-08.004.fcs | G4 | + | 3.86 | 191 | 70.1 | 5.99 | 192 | 67.9 |
5: if142014-09-08.005.fcs | G1 | - | 6.81 | 311 | 145 | 8.56 | 310 | 143 |
6: if142014-09-08.006.fcs | G2 | - | 6.37 | 337 | 153 | 7.25 | 323 | 145 |
7: if142014-09-08.007.fcs | G3 | - | 6.5 | 318 | 143 | 8.24 | 305 | 136 |
8: if142014-09-08.008.fcs | G4 | - | 6.14 | 298 | 140 | 7.66 | 288 | 131 |
9: if142014-09-08.009.fcs | C1 | + | 88.9 | 4071 | 1777 | 117 | 3978 | 1706 |
10: if142014-09-08.010.fcs | C2 | + | 83.5 | 4073 | 1547 | 120 | 3969 | 1422 |
11: if142014-09-08.011.fcs | C3 | + | 81.7 | 3957 | 1551 | 115 | 3840 | 1437 |
12: if142014-09-08.012.fcs | C4 | + | 77.2 | 3915 | 1577 | 95.7 | 3883 | 1503 |
13: if142014-09-08.013.fcs | C1 | - | 77.7 | 3686 | 1471 | 100 | 3545 | 1411 |
14: if142014-09-08.014.fcs | C2 | - | 64.5 | 3093 | 1445 | 88.3 | 2962 | 1373 |
15: if142014-09-08.015.fcs | C3 | - | 80.9 | 3971 | 1541 | 106 | 3771 | 1432 |
16: if142014-09-08.016.fcs | C4 | - | 58.9 | 2899 | 1299 | 79.7 | 2727 | 1220 |
17: if142014-09-08.017.fcs | G5 | + | 4.1 | 210 | 79.1 | 5.4 | 209 | 76.2 |
18: if142014-09-08.018.fcs | G6 | + | 4.9 | 225 | 98.2 | 7.07 | 226 | 97.6 |
19: if142014-09-08.019.fcs | G7 | + | 4.42 | 220 | 84.7 | 5.97 | 218 | 81.7 |
20: if142014-09-08.020.fcs | G8 | + | 4.19 | 211 | 81.3 | 5.68 | 213 | 79.5 |
21: if142014-09-08.021.fcs | G5 | - | 9.73 | 477 | 201 | 12.5 | 455 | 190 |
22: if142014-09-08.022.fcs | G6 | - | 9.68 | 450 | 197 | 12.9 | 449 | 196 |
23: if142014-09-08.023.fcs | G7 | - | 10.1 | 482 | 212 | 13.3 | 464 | 198 |
24: if142014-09-08.024.fcs | G8 | - | 10.1 | 489 | 216 | 13.3 | 473 | 204 |
25: if142014-09-08.025.fcs | C5 | + | 79.6 | 3958 | 1486 | 113 | 3813 | 1374 |
26: if142014-09-08.026.fcs | C6 | + | 80.8 | 4045 | 1514 | 113 | 4043 | 1425 |
27: if142014-09-08.027.fcs | C7 | + | 80.7 | 4053 | 1491 | 116 | 3921 | 1375 |
28: if142014-09-08.028.fcs | C8 | + | 79.8 | 3804 | 1555 | 103 | 3634 | 1508 |
29: if142014-09-08.029.fcs | C5 | - | 68.9 | 3268 | 1505 | 95.7 | 3128 | 1419 |
30: if142014-09-08.030.fcs | C6 | - | 80.2 | 3806 | 1639 | 110 | 3630 | 1541 |
31: if142014-09-08.031.fcs | C7 | - | 67.8 | 3240 | 1498 | 91.7 | 3041 | 1412 |
32: if142014-09-08.032.fcs | C8 | - | 83.7 | 3810 | 1675 | 107 | 3618 | 1621 |
33: if142014-09-08.033.fcs | AC1 | + | 0.895 | 40.2 | 16.8 | 0.854 | 34.9 | 12.9 |
34: if142014-09-08.034.fcs | AC3 | + | 0.539 | 28.4 | 11 | 0.785 | 27.4 | 10.7 |
35: if142014-09-08.035.fcs | AC1 | - | 0.464 | 25.8 | 9.39 | 0.605 | 25.9 | 8.65 |
36: if142014-09-08.036.fcs | AC3 | - | 0.532 | 28.3 | 10.4 | 0.661 | 26.2 | 9.23 |
- Got ratios of fluorescence in +/- aTc and am reporting them in the table below
n/a | n/a | +/- | smalls | | | bugs | |
n/a | n/a | gfpa | gfpa/fsca*1000 | gfpa/ssca*1000 | gfpa | gfpa/fsca*1000 | gfpa/ssca*1000 |
1: if142014-09-08.001.fcs | G1 | 0.771 | 0.781 | 0.772 | 0.910 | 0.797 | 0.797 |
2: if142014-09-08.002.fcs | G2 | 0.692 | 0.671 | 0.510 | 0.982 | 0.706 | 0.526 |
3: if142014-09-08.003.fcs | G3 | 0.717 | 0.726 | 0.593 | 0.947 | 0.757 | 0.613 |
4: if142014-09-08.004.fcs | G4 | 0.629 | 0.641 | 0.501 | 0.782 | 0.667 | 0.518 |
9: if142014-09-08.009.fcs | C1 | 1.144 | 1.104 | 1.208 | 1.170 | 1.122 | 1.209 |
10: if142014-09-08.010.fcs | C2 | 1.295 | 1.317 | 1.071 | 1.359 | 1.340 | 1.036 |
11: if142014-09-08.011.fcs | C3 | 1.010 | 0.996 | 1.006 | 1.085 | 1.018 | 1.003 |
12: if142014-09-08.012.fcs | C4 | 1.311 | 1.350 | 1.214 | 1.201 | 1.424 | 1.232 |
17: if142014-09-08.017.fcs | G5 | 0.421 | 0.440 | 0.394 | 0.432 | 0.459 | 0.401 |
18: if142014-09-08.018.fcs | G6 | 0.506 | 0.500 | 0.498 | 0.548 | 0.503 | 0.498 |
19: if142014-09-08.019.fcs | G7 | 0.438 | 0.456 | 0.400 | 0.449 | 0.470 | 0.413 |
20: if142014-09-08.020.fcs | G8 | 0.415 | 0.431 | 0.376 | 0.427 | 0.450 | 0.390 |
25: if142014-09-08.025.fcs | C5 | 1.155 | 1.211 | 0.987 | 1.181 | 1.219 | 0.968 |
26: if142014-09-08.026.fcs | C6 | 1.007 | 1.063 | 0.924 | 1.027 | 1.114 | 0.925 |
27: if142014-09-08.027.fcs | C7 | 1.190 | 1.251 | 0.995 | 1.265 | 1.289 | 0.974 |
28: if142014-09-08.028.fcs | C8 | 0.953 | 0.998 | 0.928 | 0.963 | 1.004 | 0.930 |
33: if142014-09-08.033.fcs | AC1 | 1.929 | 1.558 | 1.789 | 1.412 | 1.347 | 1.491 |
34: if142014-09-08.034.fcs | AC3 | 1.013 | 1.004 | 1.058 | 1.188 | 1.046 | 1.159 |
Box plot of just G1-4 and C1-4. Note that the G clones from plates (clones 5-8) have even smaller ratios.
9/9/14
Objective: assembly decoy binding site arrays
- Garima sets up a PCR and is asked to fill in notes about it.
- Results
- Charlie sets up a new set of tubes for PCR with a 5 rx master mix containing
- 190uL H2O
- 50 uL Q5 buffer
- 2.0 uL oligo 1 (gfp1-spacer-gfp1)
- 2.5 uL Q5 poly
- 5 uL dNTPs
- And then divides that in to four tubes before adding in the following volumes of oligo 2 (spacer-gfp1-space)
tube number | 1 | 2 | 3 | 4 |
uL oligo 2 | 0 | 0.5 | 1.0 | 2.0 |
- Streakiness on the row with 0.5uL of oligo 2 looks promising? IGEM protocol on the thermal cycler modified