Haynes Lab:Notebook/CRISPR Editing/2016/02/27: Difference between revisions
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Ran all cut backbones on gel, all look good! | Ran all cut backbones on gel, all look good! | ||
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Added 1ul of XbaI to MV13-KAH60 cut with SgrAI, inc at 37degC for 18 mins. Column purified, another ugly spec, maybe it's the kit? Got 48ng/ul.<br><br> | Added 1ul of XbaI to MV13-KAH60 cut with SgrAI, inc at 37degC for 18 mins. Column purified, another ugly spec, maybe it's the kit? Got 48ng/ul.<br><br> |
Revision as of 20:31, 12 March 2016
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02/27/2016Finally found the FseI in the -80! Cut 1ug of pX330A_dCas9_1x4 vector from addgene and 80ng insert (gB009) with 1ul FseI for 1h at 37degC then deactivated at 65degC for 20min. Did three tubes of the backbone.
Column purified gB014 PCR product with P169/216. Got 132ng/ul but the peak was ugly. Cut 2ug total, two reactions for 15 mins:
Column purified
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