Team:Yale/Our Project/Notebook/Week 6

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<h4> Copper Growth Assay Comparing LE392 With And Without pSB74</h4>
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<ul>
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<li>Measured the OD of the overnight LE392 cultures and found that the LE392 containing pSB74 had an OD of 0.58 while the untransformed culture had an OD of 0.87.  Wanted to let the transformed culture grow a while with IPTG prior to beginning of assay (to allow time for protein expression), but didn't want either culture to overgrow in the mean time, so diluted each as follows:<br/>
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To 1 mL of the  untransformed solution added 4 mL of plain LB, while to 3 mL of the transformed LE392 added 2 mL of LB with Amp and 10 uL of IPTG, bringing the IPTG concentration to the desired 2 mM.  Let diluted cultures grow another hour at 37˚C to reach mid-log phase. </li>
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<li> Following the second growth period, measured the OD of the untransformed culture as 0.75 while the OD of the pSB74 transformants was 0.88, but it was diluted to 0.75. </li>
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<li> Finally the solutions were loaded into the 96-well plate as follows, with each row having 12 wells and copper concentration decreasing left to right as <a href="https://2010.igem.org/Team:Yale/Our_Project/Notebook"> before. </a></li> <br/>
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Row A: Wide copper concentration range blank series with Amp & IPTG <br/>
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Row B: Narrow copper concentration range blank series with Amp & IPTG <br/>
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Row C: Wide copper concentration range transformant series with Amp & IPTG <br/>
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Row D: Narrow copper concentration range transformant series with Amp & IPTG <br/>
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Row E: Wide copper concentration range blank series<br/>
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Row F: Narrow copper concentration range blank series<br/>
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Row G: Wide copper concentration range untransformed series<br/>
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Row H: Narrow copper concentration range untransformed series<br/>
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</ul>
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<h4> Continuing ligation work </h4>
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<ul>
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<b>Miniprep and sequencing of attempt #4 transformants </b><br/>
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<li>Using the <a href="https://2010.igem.org/Team:Yale/Our_Project/Protocols/miniprep"> vacuum manifold protocol </a> miniprepped the cultures grown up from the sixteen colonies (#9-#24) that
<i>Wetlab work for this day is also recorded on pages 59 and 62-64 of the hard copy lab notebook.</i>
<i>Wetlab work for this day is also recorded on pages 59 and 62-64 of the hard copy lab notebook.</i>
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Revision as of 16:48, 27 October 2010

iGEM Yale

lab notebook: week 6 (7/12-7/18)

  • Monday 7/12--Copper growth assay of pSB74 transformants & continued ligation work
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  • Tuesday 7/13--
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  • Wednesday 7/14--
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  • Thursday 7/15--
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  • Friday 7/16--
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Retrieved from "http://2010.igem.org/Team:Yale/Our_Project/Notebook/Week_6"