https://eln.excellgen.com/index.php?action=history&feed=atom&title=3244_pERD-sfGFP 3244 pERD-sfGFP - Revision history 2026-10-18T07:07:37Z Revision history for this page on the wiki MediaWiki 1.44.0 https://eln.excellgen.com/index.php?title=3244_pERD-sfGFP&diff=18930&oldid=prev Crm at 16:48, 7 August 2018 2018-08-07T16:48:05Z <p></p> <p><b>New page</b></p><div>== &#039;&#039;&#039;Cloning of 3244 pERD-sfGFP&#039;&#039;&#039; ==<br /> <br /> &#039;&#039;&#039;Purpose:&#039;&#039;&#039; Cloning of 3244 pERD-sfGFP<br /> <br /> &#039;&#039;&#039;Date: &#039;&#039;&#039;07, 25, 2018<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: 3244&#039;&#039;&#039;<br /> <br /> &#039;&#039;&#039;Experimenters: &#039;&#039;&#039;yijia<br /> <br /> == &#039;&#039;&#039;Results&#039;&#039;&#039; ==<br /> <br /> === Cloning ===<br /> <br /> PCR from 128 pET30b-sfGFP<br /> <br /> &#039;&#039;&#039;PCR of 3244&#039;&#039;&#039;<br /> <br /> &#039;&#039;&#039;Size:780bp&#039;&#039;&#039;<br /> <br /> &#039;&#039;&#039;method:&#039;&#039;&#039;<br /> <br /> first: 1 ul of 1 uM F3--R2, 10 cycles;<br /> <br /> 2nd: 1 ul of 1 uM F2--R2, 10 cycles;<br /> <br /> 3nd 1ul of 10 uM 3244-F1--3244-R1 25 cycles;<br /> <br /> clone into 3242 pERD using Nde1-HindIII;<br /> <br /> &#039;&#039;&#039;3244 PCR&#039;&#039;&#039;<br /> <br /> [[File:IMG 1225.JPG|RTENOTITLE]]<br /> <br /> &#039;&#039;&#039;Vector digestion&#039;&#039;&#039;<br /> <br /> Vector is &#039;&#039;&#039;3242&#039;&#039;&#039;<br /> <br /> buffer:Fastdigest<br /> <br /> ENzymes:Nde1+HindIII<br /> <br /> Time:1hour<br /> <br /> [[File:IMG 1197.JPG|RTENOTITLE]]<br /> [[Category:Cloning]]<br /> <br /> === &amp;nbsp;Colony screen ===<br /> <br /> &#039;&#039;&#039;3244 PCR screening&#039;&#039;&#039;<br /> <br /> [[File:IMG 1245.JPG|RTENOTITLE]]<br /> <br /> &#039;&#039;&#039;Miniprep digestion998_129_3244&#039;&#039;&#039;<br /> <br /> [[File:IMG 1250.JPG|RTENOTITLE]]<br /> <br /> &#039;&#039;&#039;Miniprep digestion 2_3244_129_998&#039;&#039;&#039;<br /> <br /> [[File:IMG 1252.JPG|RTENOTITLE]]<br /> <br /> == &#039;&#039;&#039;Result intepretation/Summary&#039;&#039;&#039; ==<br /> <br /> == &#039;&#039;&#039;Conclusion&#039;&#039;&#039; ==</div> Crm