Haynes Lab:Notebook/CRISPR Editing/2014/07/07

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07/07/2014

Miniprepped the 6 colonies I picked yesterday from ligation pX330 - g14.

Sample Read# 260 280 260/280 ng/┬ÁL
Colony 12.10E-020.012.11521.036
Colony 21.70E-020.011.71716.52
Colony 31.90E-029.00E-032.10518.534
Colony 42.60E-021.50E-021.76425.633
Colony 52.20E-021.40E-021.62421.991
Colony 66.90E-023.70E-021.86669.266



Cut mini preps with EcoRI and BsbI. Proper insertion of the gRNA will destroy the BsbI sites. If the colony is no cut or single cut backbone religating, I'll get bands of 5.2kb and 3.2kb. If the insert is there, I'll get one band of 8.4kb.

' Volume
EcoRI0.5
BsbI0.5
10X ligation buffer2
DNA17



Ran on gel, didn't see anything. Just a faint smear for colony 6.



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