https://eln.excellgen.com/index.php?action=history&feed=atom&title=1727_pT7A30
1727 pT7A30 - Revision history
2026-10-18T08:06:59Z
Revision history for this page on the wiki
MediaWiki 1.44.0
https://eln.excellgen.com/index.php?title=1727_pT7A30&diff=17276&oldid=prev
Crm at 23:00, 13 March 2018
2018-03-13T23:00:27Z
<p></p>
<p><b>New page</b></p><div>== '''Cloning of 1727 pT7A30''' ==<br />
<br />
'''Purpose:''' Cloning of 1727 pT7A30 <br />
<br />
'''Date: '''2013-5-6 <br />
<br />
'''Notebook:0''' <br />
<br />
'''Page:''' <br />
<br />
'''Clone ID:1727''' <br />
<br />
'''Experimenters:''' shanyi <br />
<br />
== '''Results''' ==<br />
<br />
=== cloning ===<br />
<br />
digest pT7A3 vector or any pT3A3 clone with Xba1 + Xho1---> cut vector out<br/>digest pET30b with Xba1 + Xho1--> cut out 232 bp MCS fragment<br />
<br />
[[File:Digest for 1727 05 06 2013.JPG|RTENOTITLE]]<br />
<br />
ligate and transform<br />
<br />
=== Colony Screening ===<br />
<br />
Date: 2013-3-<br />
<br />
Make miniprep and digest with Kpn1 + Sma1, should get one band (the parenta vector pET28b does not have Kpn1 site, and there is one Kpn1 site in pET30b)<br />
<br />
[[File:1727DIGEST.JPG|RTENOTITLE]]<br />
<br />
1727digest_with_XbaIand_XhoI<br />
<br />
[[File:1727digest with XbaIand XhoI.JPG|RTENOTITLE]]<br />
<br />
'''Miniprep of 945pMiniT2_947pMiniT2_3131_1727_1834'''<br />
<br />
945pMiniT2:1-4 EcoRI<br />
<br />
947pMiniT2:5-8 n EcoRI<br />
<br />
3131:9-13 BssHII+XhoI<br />
<br />
1727:14 XbaI+NcoI<br />
<br />
1834:15 XbaI+NcoI<br />
<br />
Buffer:Fastdigest<br />
<br />
Time:30Min<br />
<br />
'''Results:945pMiniT2 and 947pMiniT2 are good, so sent them to do sequencing. 3131 is not good, 1727 has no band, 1834 has one band.'''<br />
<br />
[[File:Miniprep of 945 947 1727 1834.JPG|RTENOTITLE]]<br />
<br />
== '''Result intepretation/Summary''' ==<br />
<br />
<br> <br />
<br />
== '''Conclusion''' ==<br />
<br />
<br> <br />
<br />
<br> <br />
<br />
[[Category:Cloning]]</div>
Crm