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3154 pCMV5-HA-VH-mKappa - Revision history
2026-10-18T10:47:53Z
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Crm at 23:19, 13 March 2018
2018-03-13T23:19:44Z
<p></p>
<p><b>New page</b></p><div>2/17/2018<br />
<br />
subcloning<br />
<br />
1: insert,&nbsp;<br />
<br />
2: vector, top band<br />
<br />
[[File:3154 subcloning.JPG|RTENOTITLE]]<br />
<br />
miniprep digestion<br />
<br />
[[File:3154 digestion.JPG|RTENOTITLE]]<br />
<br />
BamH1 digestion, to see if the clone has only one BamH1 site<br />
<br />
[[File:3154 BamH1 digestionJPG.jpeg|RTENOTITLE]]<br />
<br />
<br />
<br />
[[File:3154 Kpn1-Not1.JPG|RTENOTITLE]]<br />
<br />
<br />
<br />
[[File:3154 Xho1.JPG|RTENOTITLE]]<br />
<br />
== Redo ==<br />
<br />
[[File:2493 3129 digestion1.JPG|RTENOTITLE]]<br />
<br />
[[File:2493 3129 digestion2.JPG|RTENOTITLE]]<br />
<br />
2/19/2018<br />
<br />
Subclone HA heavy 3029-1 in pMiniT2&nbsp;to 3130 pCMV5-HA-VL-mIgGkappa<br />
<br />
to create an intermediate cloning vector which can be used for subcloning of Heavy chain&nbsp;<br />
<br />
because there is an Bamh1 site in the IgG2b constant region[[File:3130 3129 digestion 2 19 18.JPG|RTENOTITLE]]<br />
<br />
<br />
Miniprep digestion for 3154_3129<br />
<br />
Digestion buffer:Fastdigest<br />
<br />
Restriction enzymes:<br />
<br />
3154:BamH I+ Kpn I'''insert length:527'''<br />
<br />
3129:Kpn I+Not I'''inset length:1034'''<br />
<br />
[[File:Miniprep digestion for 3154 3129.JPG|RTENOTITLE]]<br />
<br />
Result:seems good, chose 3129-1 to do sequencing<br />
[[Category:Cloning]]</div>
Crm