Haynes Lab:Notebook/CRISPR Editing/2014/09/21: Difference between revisions
Rene M Davis (talk | contribs) |
Rene M Davis (talk | contribs) |
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add 5 ul of MM 2 to each tube | add 5 ul of MM 2 to each tube | ||
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PCR | PCR <br> | ||
98°C for 3 min | :98°C for 3 min | ||
:98°C for 10s | :36 cycles | ||
:66°C for 30s | ::98°C for 10s | ||
:72°C for 60s | ::66°C for 30s | ||
: | ::72°C for 60s | ||
72°C for 10 min | :72°C for 10 min | ||
4°C forever | :4°C forever | ||
<br><br> | <br><br> | ||
transferred 40ul of the first three annealing reactions and 35ul of the last two to new tubes. added 1ul enhancer and 2ul of surveyor nuclease. incubate at 42deg C for 1 hour. | transferred 40ul of the first three annealing reactions and 35ul of the last two to new tubes. added 1ul enhancer and 2ul of surveyor nuclease. incubate at 42deg C for 1 hour. | ||
Revision as of 15:03, 21 September 2014
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09/21/2014Ran the PCR for gRNA 20 samples from yesterday on a gel. Ran 3μL of each sample and 2μL and 1μL of ladder. Only the first three worked, even though the other two worked the other day.
Master mix 1
add 41ul of MM 1 to each tube
add 5 ul of MM 2 to each tube
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