https://eln.excellgen.com/index.php?action=history&feed=atom&title=1298_pET-6H-MPGa
1298 pET-6H-MPGa - Revision history
2026-10-18T02:17:41Z
Revision history for this page on the wiki
MediaWiki 1.44.0
https://eln.excellgen.com/index.php?title=1298_pET-6H-MPGa&diff=7019&oldid=prev
Paul.xu at 18:49, 14 November 2012
2012-11-14T18:49:02Z
<p></p>
<p><b>New page</b></p><div>== '''Cloning of 1298 pET-6H-MPGa''' ==<br />
<br />
'''Purpose:''' Cloning of 1298 pET-6H-MPGa <br />
<br />
'''Date: '''August 13, 2012<br> <br />
<br />
'''Notebook:&nbsp;''' <br />
<br />
'''Page:''' <br />
<br />
'''Clone ID: '''1298 <br />
<br />
'''Experimenters: '''Jennifer Truong <br />
<br />
== '''Results''' ==<br />
<br />
=== Cloning<br> ===<br />
<br />
Date:&nbsp;August 13, 2012 <br />
<br />
Cloning Method <br />
<br />
PCR gene synthesis, see lane 3, expected size 114 bp <br />
<br />
[[Image:1298 0813 Amp.JPG]] <br />
<br />
Cut out and purify DNA<br>Digest with Nco1 + Nde1, ligate to Nco1 + Nde1 digested 1128 pDB.His.PfuMBP-(tev) overnight, transform <br />
<br />
<br> Date:&nbsp;August 14, 2012 <br />
<br />
Insert disappeared after digestion, need to reamplify, see lane 2 <br />
<br />
[[Image:0814 Amp.JPG]] <br />
<br />
<br> <br />
<br />
Date:&nbsp;August 15, 2012<br> <br />
<br />
Digest with Nco1 + Nde1 (lane 3), ligate to Nco1 + Nde1 digested pDB.His.MBP, V14 (lane 4), transform <br> <br />
<br />
[[Image:1298 0815 Digestion.JPG]] <br />
<br />
Date:&nbsp;August 17, 2012<br> <br />
<br />
[[Image:1298 0817 Amp.JPG]]<br> <br />
<br />
Digest with Nco1 + Nde1 (lane 2), ligate to Nco1 + Nde1 digested pDB.His.MBP, V14 , transform <br> <br />
<br />
[[Image:1298 Insert Digestion.JPG]]<br> <br />
<br />
=== <u></u>Colony Screening<br> ===<br />
<br />
Date:&nbsp;August 20, 2012<br> <br />
<br />
[[Image:1298 Screening.JPG]] <br />
<br />
No inserts <br />
<br />
== '''Result intepretation/Summary''' ==<br />
<br />
<br> <br />
<br />
== '''Conclusion''' ==<br />
<br />
<br> <br />
<br />
<br> <br />
<br />
== '''Cloning of 1298 pET-6H-MPGa''' ==<br />
<br />
'''Purpose:''' Cloning of 1298 pET-6H-MPGa <br />
<br />
'''Date: '''August 30, 2012<br> <br />
<br />
'''Notebook:&nbsp;''' <br />
<br />
'''Page:''' <br />
<br />
'''Clone ID: '''1298 <br />
<br />
'''Experimenters: '''yingying lu <br />
<br />
== '''Results''' ==<br />
<br />
=== Cloning<br> ===<br />
<br />
Date:&nbsp;August 30, 2012 <br />
<br />
Cloning Method <br />
<br />
PCR gene synthesis, see lane 3, expected size 114 bp <br />
<br />
[[Image:1298+1348p1+1350 PCR 8-31-2012.JPG]]<br> <br />
<br />
[[Image:1298+1324+1346+1347+1350+1356 clone 8-31-2012.JPG]] <br />
<br />
Cut out and purify DNA<br>Digest with Nco1 + Nde1, ligate to Nco1 + Nde1 digested V14 pDB.His-MBP overnight, transform <br />
<br />
<br> <br />
<br />
<br> <br />
<br />
=== <u></u>Colony Screening<br> ===<br />
<br />
Date: 2012-9-4<br> <br />
<br />
no insert <br />
<br />
== '''Result intepretation/Summary''' ==<br />
<br />
<br> <br />
<br />
== '''Conclusion''' ==<br />
<br />
<br> <br />
<br />
<br> <br />
<br />
== Cloning of 1298 pET-6H-MPGa ==<br />
<br />
'''Purpose:''' Cloning of 1298 pET-6H-MPGa <br />
<br />
'''Date: '''2012-10-19---2012-10-23<br> <br />
<br />
'''Notebook:&nbsp;''' <br />
<br />
'''Page:''' <br />
<br />
'''Clone ID: '''1298 <br />
<br />
'''Experimenters:&nbsp; '''shanyi&amp; yingying<br> <br />
<br />
== '''Results''' ==<br />
<br />
=== Cloning<br> ===<br />
<br />
[[Image:1298pcr.JPG]]<br />
<br />
<br> <br />
<br />
<u></u>Colony Screening<br> <br />
<br />
Date: 2012-10-22<br> <br />
<br />
5ul sample <br />
<br />
[[Image:1298+1308+1309+1310 digest detect 10-22-2012.JPG]]<br> <br />
<br />
2012-10-23 <br />
<br />
10ul sample <br />
<br />
[[Image:1298 digest detect 10-23-2012.JPG]] <br />
<br />
== '''Result intepretation/Summary''' ==<br />
<br />
<br> <br />
<br />
== '''Conclusion''' ==<br />
<br />
<br> <br />
<br />
[[Category:Cloning]]</div>
Paul.xu