Haynes Lab:Notebook/CRISPR Editing/2015/01/31

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01/30/2015

miniprepped colonies from topo cloning plate for g034

Sample Read# 260 280 260/280 ng/┬ÁL
Topo clone 10.020.0111.82719.983
Topo clone 20.0570.031.88257.143
Topo clone 30.5820.3021.924581.887
Topo clone 40.0180.0082.33317.68
Topo clone 50.0460.0251.85146.218
Topo clone 60.040.0211.89640.046


Ran gradient PCR reactions from yesterday on gel. Setting AT of 66 as 100%, estimated the drop in efficiency. Def not quantitative. There was an obvious off target band around 800bp that faded and disappeared at higher annealing temperatures.

Annealing Temps Efficiency off target band ~800bp
66100yes
67100yes
68100yes
6980faint to gone
7080gone
7260gone


Split Cameron's cells into two T-25 flasks.



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