https://eln.excellgen.com/index.php?action=history&feed=atom&title=1255_pTXB1-C6H2 2026-10-18T05:43:20Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1255_pTXB1-C6H2&diff=5009&oldid=prev 2012-07-09T15:58:26Z

<p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1255 pTXB1-C6H2&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1255 pTXB1-C6H2 <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;July 6, 2012&lt;br&gt; <br /> <br /> &#039;&#039;&#039;Notebook:&amp;nbsp;&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID: &#039;&#039;&#039;1255 <br /> <br /> &#039;&#039;&#039;Experimenters: &#039;&#039;&#039;Jennifer Truong <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === Cloning ===<br /> <br /> Amplified from 1126 (pTXB1) template with pTXB1-Xho1-F and pTXB1-Pst-R2 primers <br /> <br /> Cut out band in lane 4, expected size 624 bp, purified and digested&amp;nbsp; with restriction enzymes Xho1 and Pst1 <br /> <br /> [[Image:0706 amp.JPG]]&lt;br&gt; <br /> <br /> Digested DNA and vector with restriction enzymes Xho1 and Pst1 &lt;br&gt; <br /> <br /> [[Image:0706 digestion.JPG]] <br /> <br /> Cut out bands from lane 3 and 4, purify, ligate and transform, plate and let grow overnight&lt;br&gt; <br /> <br /> Is lane 4 vector pTXB1? it should have two bands, one vector, one original insert.&lt;br&gt; <br /> <br /> Dropped DNA band, and vector band is a little too light, reamplified&lt;br&gt; <br /> <br /> [[Image:1255 amp.JPG]]&lt;br&gt; <br /> <br /> Cut out lane 1 (using Phusion from Thermo) and 2 (using homemade Phusion), expected size 624 bp, purified and digested&amp;nbsp; with restriction enzymes Xho1 and Pst1, &lt;br&gt; <br /> <br /> [[Image:0707_digestion.JPG]]<br /> <br /> Lane 1 and 2 are the digested DNA and vector with Xho1+ Pst1 (from Phusion from Thermo)<br /> <br /> Lane 3 and 4 are the digested DNA&amp;nbsp;and vector with Xho1 and Pst1 (from Homemade Phusion)<br /> <br /> Lane 4 digestion does not look good (vector)&amp;nbsp;&lt;br&gt; <br /> <br /> Only cut out lane 1 and 2, purified and ligated together overnight. Paul transformed with Top10 cells and plated<br /> <br /> === Colony Screening&lt;br&gt; ===<br /> <br /> Date:&amp;nbsp;June 9, 2012<br /> <br /> [[Image:0709_screening.JPG]]<br /> <br /> Digested MiniPrep plasmids with restriction enzymes Xho1 and Pst1<br /> <br /> May have postive insert for all samples, but unclear<br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039;&lt;br&gt; ==<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039;&lt;br&gt; ==<br /> <br /> [[Category:Cloning]]</div>

Xiaojing.Wang