https://eln.excellgen.com/index.php?action=history&feed=atom&title=1601_pT7A3-6His-GST-i3C
1601 pT7A3-6His-GST-i3C - Revision history
2026-10-18T14:13:34Z
Revision history for this page on the wiki
MediaWiki 1.44.0
https://eln.excellgen.com/index.php?title=1601_pT7A3-6His-GST-i3C&diff=8233&oldid=prev
Paul.xu at 20:08, 14 February 2013
2013-02-14T20:08:32Z
<p></p>
<p><b>New page</b></p><div>== '''Cloning of 1601 pT7A3-6His-GST-i3C''' ==<br />
<br />
'''Purpose:''' Cloning of 1601 pT7A3-6His-GST-i3C <br />
<br />
'''Date: '''2013-1-28---2013-1-30 <br />
<br />
'''Notebook:0''' <br />
<br />
'''Page:''' <br />
<br />
'''Clone ID:1601''' <br />
<br />
'''Experimenters:''' Shanyi <br />
<br />
== '''Results''' ==<br />
<br />
=== subcloning ===<br />
<br />
subclone i3C from 1554 pT7A3-MBP-i3C to 1597 pT7A3-6XHis.GST using Nde1 + Xho1<br> <br />
<br />
{| cellspacing="1" cellpadding="1" border="1" style="width: 281px; height: 66px;"<br />
|-<br />
| <br> <br />
| band 1<br> <br />
| band 2<br><br />
|-<br />
| <br> <br />
| 1597 pT7A3-6XHis.GST<br> <br />
| 1554 pT7A3-MBP-i3C<br><br />
|-<br />
| enzymes<br> <br />
| colspan="2" | &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Nde1 + Xho1<br><br />
|}<br />
<br />
[[Image:DIGEST FOR 1601 1 25 2013.JPG]] <br />
<br />
=== Colony Screening ===<br />
<br />
Date: 2013-1-30 <br />
<br />
Make miniprep and digest with Nde1 + Xho1. <br />
<br />
[[Image:1503 1601 1552 1122 1123digest.JPG]] <br />
<br />
digest positive clone with Nco1 alone to confirm GST not MBP is in the construct. Nco1 will cut out one band from MBP<br><br> <br />
<br />
[[Image:1601digest with NcoI.JPG]]<br> <br />
<br />
<br> <br />
<br />
<br />
<br />
== '''Conclusion''' ==<br />
<br />
== Protein Expression ==<br />
<br />
<br />
[[1625 pCold-GST-(3c)-KEAP1#Protein_Expression|1625_pCold-GST-(3c)-KEAP1#Protein_Expression]] <br />
<br />
[[Category:Cloning]]</div>
Paul.xu