https://eln.excellgen.com/index.php?action=history&feed=atom&title=2214_pET30b-Dest-Avi-8H-BirA 2214 pET30b-Dest-Avi-8H-BirA - Revision history 2026-10-18T16:06:54Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=2214_pET30b-Dest-Avi-8H-BirA&diff=12958&oldid=prev Excellgen at 16:41, 26 August 2014 2014-08-26T16:41:15Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 2214 pET30b-Dest-Avi-8H-BirA&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of2214 pET30b-Dest-Avi-8H-BirA&#039;&#039;&#039;==&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;2014-8---- <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:2214&#039;&#039;&#039; <br /> <br /> &#039;&#039;&#039;Experimenters:&#039;&#039;&#039;&amp;nbsp; shanyi <br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === CLONING ===<br /> <br /> PCR from BL21 genomic DNA using these 3 oligos: <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt;1ul of 10 uM of BirA-HindIII-F1 (8_21_2013 A03)&lt;br&gt;atcgacAAGCTTGCGGCCGCActcgagggactgaacgatatttttgaggcgcaaaaaattgagtggcatgaaCAC <br /> <br /> 1ul of 1 uM of BirA-F2 (8_21_2013 A04)&lt;br&gt;attgagtggcatgaaCACCACCACCACCACCACTAACTTTAAGAAGGAGTCTCTCCCATGAAGGATAACACCGTGCCAC <br /> <br /> &lt;br&gt;1ul of 10 uM of BirA-Sal1-R (8_21_2013 A05)&lt;br&gt;(CTGCGTAGTGCAGAAAAATAA gtcgac cgatca)&lt;br&gt;tgatcggtcgacTTATTTTTCTGCACTACGCAG <br /> <br /> [[Image:2214 2238 PCR.JPG]]&lt;br&gt; <br /> <br /> cut 1 kb PCR product with HindIII + Sal1, &lt;br&gt;clone into HindIII + Xho1 digested 2213 pET30b-Dest <br /> <br /> &lt;br&gt; <br /> <br /> [[Image:Digest for 2300 2214 2198 212.JPG]]&lt;br&gt;&lt;br&gt; <br /> <br /> === Colony Screening ===<br /> <br /> Date: 2014_8_ <br /> <br /> Make miniprep, use EagI to find positive clone&lt;br&gt; <br /> <br /> 22[[Image:2214_2232_2234digest.JPG]]<br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; <br /> <br /> &lt;br&gt; &lt;br&gt; <br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> *&lt;br&gt;<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==<br /> <br /> [[Category:Cloning]]</div> Excellgen