https://eln.excellgen.com/index.php?action=history&feed=atom&title=1265_pET30b-Tks 2026-10-18T03:32:34Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=1265_pET30b-Tks&diff=5444&oldid=prev 2012-08-03T17:59:22Z

<p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 1265 pET30b-Tks&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1265 pET30b-Tks <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;July 13, 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;1265 <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 /> Date:&amp;nbsp;July 13, 2012 <br /> <br /> [[Image:1265 0713 amp.JPG]]&lt;br&gt; <br /> <br /> Gene synthesis of 4 fragments and then join by PCR, expected size 2.4 kb&lt;br&gt; <br /> <br /> Not correct size, need to reamplify on Monday&lt;br&gt; <br /> <br /> Procedure:&amp;nbsp;4 PCR&amp;nbsp;reactions for each fragment, (1 uL 10 uM&amp;nbsp;mix, 1 uL&amp;nbsp;for template) with PHUSION&amp;nbsp;enzyme, 10 cycles. Then took 6 uL&amp;nbsp;from each fragment, added 1 uL&amp;nbsp;more of PHUSION&amp;nbsp;enzyme and 1 uL of Tks-1-1 and Tks3-18 (primers), PCR for 20 more cycles with 2:30 minutes extension time&lt;br&gt; <br /> <br /> Looks like fragments did not join together? Band is around ~600 bp, which is the size of each fragment... <br /> <br /> === Colony Screening&lt;br&gt; ===<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 /> ==<br /> <br /> == &#039;&#039;&#039;Cloning of 1265-pET30b-Tks&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 1025-pET28b-SM2 <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2012-7-31---2012-8-2<br /> <br /> &#039;&#039;&#039;Notebook:0&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Page:&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Clone ID:1265&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039; yingying <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === cloning ===<br /> <br /> [[Image:1265 PCR1 7-31-2012.JPG]]&lt;br&gt; <br /> <br /> Gene synthesis of 4 fragments&lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:1264+1265 PCR 7-31-2012.JPG]]&lt;br&gt; <br /> <br /> 2nd PCR join four fragement together by PCR method&lt;br&gt; <br /> <br /> {| cellspacing=&quot;1&quot; cellpadding=&quot;1&quot; border=&quot;1&quot;<br /> |-<br /> | <br /> |}<br /> <br /> === Colony Screening ===<br /> <br /> Date: 2012-8-2<br /> <br /> Make miniprep and digest with <br /> <br /> no insert&lt;br&gt; &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> *<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> {|<br /> |-<br /> | &lt;br&gt;<br /> |}<br /> <br /> [[Category:Cloning]]</div>

Paul.xu