Haynes Lab:Notebook/CRISPR Editing/2014/09/24: Difference between revisions
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Rene M Davis (talk | contribs) (Autocreate 2014/09/24 Entry for Haynes_Lab:Notebook/CRISPR_Editing) |
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== | ==09/24/2014== | ||
Ran 4x50ul PCR reactions of gRNA 20 and gRNA 29 +dox samples 1 and 2 on a gel. All had one band, mostly high yields, one of gRNA29 sample 1 had a lower yield. | |||
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Set up four sets of annealing reactions for Luc14 samples, only annealed two of them, then cut all of them. One set was 0.5ul of Nuclease for 1hour, other set was 1ul for 30 minutes (paused the other one at about 28 minutes left in the incubation. This will tell me if nuclease is randomly cutting (if it cuts PCR product that wasn't annealed) and if reducing the time or the amount of nuclease will get rid of the smear I saw yesterday. | |||
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Purify the gRNA 20 and 29 +dox samples using genelute PCR kit. Elute in 50ul elution solution. | |||
Revision as of 10:47, 24 September 2014
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09/24/2014Ran 4x50ul PCR reactions of gRNA 20 and gRNA 29 +dox samples 1 and 2 on a gel. All had one band, mostly high yields, one of gRNA29 sample 1 had a lower yield.
Set up four sets of annealing reactions for Luc14 samples, only annealed two of them, then cut all of them. One set was 0.5ul of Nuclease for 1hour, other set was 1ul for 30 minutes (paused the other one at about 28 minutes left in the incubation. This will tell me if nuclease is randomly cutting (if it cuts PCR product that wasn't annealed) and if reducing the time or the amount of nuclease will get rid of the smear I saw yesterday.
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