Haynes Lab:Notebook/CRISPR Editing/2015/01/31: Difference between revisions

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Ran gradient PCR reactions from yesterday on gel.
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.
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Temps'''
| align="center" style="background:#f0f0f0;"|'''Efficiency'''
| align="center" style="background:#f0f0f0;"|'''off target band ~800bp'''
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| 66||100||yes
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| 67||100||yes
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| 68||100||yes
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| 69||80||faint to gone
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| 70||80||gone
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| 72||60||gone
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Split Cameron's cells into two T-25 flasks.
Split Cameron's cells into two T-25 flasks.

Revision as of 11:28, 3 February 2015

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01/30/2015

miniprepped colonies from topo cloning plate for g034

Sample Read# 260 280 260/280 ng/µL
Topo clone 1 0.02 0.011 1.827 19.983
Topo clone 2 0.057 0.03 1.882 57.143
Topo clone 3 0.582 0.302 1.924 581.887
Topo clone 4 0.018 0.008 2.333 17.68
Topo clone 5 0.046 0.025 1.851 46.218
Topo clone 6 0.04 0.021 1.896 40.046


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.

Temps Efficiency off target band ~800bp
66 100 yes
67 100 yes
68 100 yes
69 80 faint to gone
70 80 gone
72 60 gone


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



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