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/µL
Colony 1 2.10E-02 0.01 2.115 21.036
Colony 2 1.70E-02 0.01 1.717 16.52
Colony 3 1.90E-02 9.00E-03 2.105 18.534
Colony 4 2.60E-02 1.50E-02 1.764 25.633
Colony 5 2.20E-02 1.40E-02 1.624 21.991
Colony 6 6.90E-02 3.70E-02 1.866 69.266



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
EcoRI 0.5
BsbI 0.5
10X ligation buffer 2
DNA 17



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