https://eln.excellgen.com/index.php?action=history&feed=atom&title=PLenti-IRES-TFR-EGFP 2026-10-18T11:50:55Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=PLenti-IRES-TFR-EGFP&diff=1315&oldid=prev 2011-07-17T20:00:42Z

<p>moved <a href="/index.php?title=PLenti-IRES-TFR-EGFP%27" class="mw-redirect" title="PLenti-IRES-TFR-EGFP&#039;">PLenti-IRES-TFR-EGFP&#039;</a> to <a href="/index.php?title=PLenti-IRES-TFR-EGFP" title="PLenti-IRES-TFR-EGFP">PLenti-IRES-TFR-EGFP</a></p> <p><b>New page</b></p><div>CloneID: 252 <br /> <br /> Date: 2011-6-24to 2011-6-28&lt;br&gt;<br /> <br /> &lt;br&gt;<br /> <br /> &#039;&#039;&#039;PCR, DIGESTION, GEL&amp;nbsp;ELECTROPHORESIS:&#039;&#039;&#039;&lt;br&gt; <br /> <br /> PCR1) 1ul F1, 1ul&amp;nbsp;P8, 2ul primer mix, 5ul 5x buffer, 0.5ul 50x dNTP, 14.5ul H2O, 1ul pfu enzyme&lt;br&gt;<br /> <br /> PCR2) 1ul F2, 1ulR, 1ul EGFP template, 5ul 5x buffer, 0.5ul dNTP, 15.5ul H2O, 1ul pfu enzyme&lt;br&gt;<br /> <br /> PCR3) 5 ul PCR1, 5ul PCR2, 1ul F1, 1ul&amp;nbsp;R, 8ul 5x buffer, 1ul dNTP, 2ul pfu enzyme, 27ul H2O&lt;br&gt;<br /> <br /> &lt;br&gt;<br /> <br /> Used MluI+XhoI enzymes to digest, 100% in buffer 3 <br /> <br /> 1)&amp;nbsp;pLentiplus vector:&amp;nbsp;5ul pLentiplus, 0.4ul Xho1, 0.4ul Mlu1, 1.5ul buffer 3, 7.7ul H2O&lt;br&gt;<br /> <br /> 2) PCR3 product:&amp;nbsp;12.7ul purified DNA, 0.4ul Xho1, 0.4ul Mlu1, 1.5ul buffer 3 (75ºC for ~15 min after digestion complete)&lt;br&gt;<br /> <br /> &lt;br&gt;<br /> <br /> Ran agarose gel for pLentiplus vector and PCR3 product&lt;br&gt;<br /> <br /> (Image to be uploaded later)&lt;br&gt;<br /> <br /> Lanes: <br /> <br /> &lt;br&gt;<br /> <br /> &#039;&#039;&#039;COLONY&amp;nbsp;SCREENING:&#039;&#039;&#039;&lt;br&gt;<br /> <br /> PCR:&lt;br&gt;<br /> <br /> 11 colonies -&lt;br&gt;<br /> <br /> each contains:&amp;nbsp;1ul template (H2O+colony), 1ul A3, 1ul B12, 2.5ul 10x buffer, 0.8ul Taq enzyme, 0.2ul dNTP, 18.5ul H2O&lt;br&gt;<br /> <br /> <br /> [[Image:PLenti-IRES-TFR-EGFP-ColonyScreening.jpg]] &lt;br&gt;<br /> <br /> Lanes:&lt;br&gt;<br /> <br /> Marker&lt;br&gt;<br /> <br /> 1: Marker<br /> <br /> 2: colony 1&lt;br&gt;<br /> <br /> 3: colony 2&lt;br&gt;<br /> <br /> 4: colony 3&lt;br&gt;<br /> <br /> 5: colony 4&lt;br&gt;<br /> <br /> 6: colony 5&lt;br&gt;<br /> <br /> 7: colony 6&lt;br&gt;<br /> <br /> 8: colony 7&lt;br&gt;<br /> <br /> 9: colony 8&lt;br&gt;<br /> <br /> 10: colony 9&lt;br&gt;<br /> <br /> 11: colony 10&lt;br&gt;<br /> <br /> 12: colony 11&lt;br&gt;<br /> <br /> Took DNA from colonies 5-8 and 10-11<br /> <br /> (pg. 001143)&lt;br&gt;<br /> <br /> &lt;br&gt;<br /> <br /> &lt;br&gt;<br /> <br /> &lt;br&gt;<br /> <br /> &lt;br&gt;<br /> <br /> [[Category:Cloning|Cloning]]</div>

Paul.xu