Haynes Lab:Notebook/CRISPR Editing/2015/03/19: Difference between revisions

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(Autocreate 2015/03/19 Entry for Haynes_Lab:Notebook/CRISPR_Editing)
 
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==03//2015==
==03/19/2015==
Final Surveyor test.
:Sample: Luc14 cells treated with g031 rep 1 <br>
:PCR product: nested PCR P197/198, P149/174<br>
:Known editing efficiency: ~40%.
:Expected band sizes: 337, 210, 127
<br>
Will change surveyor enzyme volume (top row) and incubation time (left column), run 11 different variations so I can use one chip:
 
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<br>
Annealed 400ng in 15ul with 1ul of 10x hybridization buffer (should have used 1.5ul). Ran on new PCR machine with recommended annealing protocol
<br>




Revision as of 11:42, 19 March 2015

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03/19/2015

Final Surveyor test.

Sample: Luc14 cells treated with g031 rep 1
PCR product: nested PCR P197/198, P149/174
Known editing efficiency: ~40%.
Expected band sizes: 337, 210, 127


Will change surveyor enzyme volume (top row) and incubation time (left column), run 11 different variations so I can use one chip:

0.25 0.5 0.75
15 x x x
20 x x x
30 x x x
45 x x


Annealed 400ng in 15ul with 1ul of 10x hybridization buffer (should have used 1.5ul). Ran on new PCR machine with recommended annealing protocol



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