Haynes Lab:Notebook/CRISPR Editing/2014/09/22: Difference between revisions
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Ran dilutions of ladder on gel with tiniest comb size to determine detection threshold using Sybr and our new gel imager. | Ran dilutions of ladder on gel with tiniest comb size to determine detection threshold using Sybr and our new gel imager. | ||
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Set up 4-50ul PCR rxns for each Luc14 samples (20 total). Made one master mix of water, buffer, dNTPs, and primers, split into 5 tubes, added template, DMSO, and Phusion, then split each of those 5 into 4 tubes. Ran overnight. | |||
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Revision as of 18:16, 23 September 2014
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09/22/2014Cut the annealed Luc14 reactions, but not gRNA 21 because that one didn't amplify. added 40ul of annealed reaction to a new PCR tube, added 1ul of enhancer and 2ul of Surveyor nuclease. Incubated at 42°C for one hour, kept on ice after. Forgot to add stop solution. ran gel after about an hour or so on ice.
add 31ul of MM 1 to each tube
add 15 ul of MM 2 to each tube
Ran dilutions of ladder on gel with tiniest comb size to determine detection threshold using Sybr and our new gel imager.
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