Haynes Lab:Notebook/CRISPR Editing/2015/01/07

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01/07/2015

Set up 2-50μL PCR reactions for untreated Luc14 gDNA and a subset of treated Luc14.

sample
Untreated
23
25
29
30
32
33
34
36
39
43
44
45
46


Mastermix 1 ' '
1rxn 28.3rxns
H20 30 849
HF 5X MasterMix 10 283
dNTPs 1 28.3
FP 2.5 70.75
RP 2.5 70.75
46 1301.8
Mastermix 1
1rxn 2.1rxns
MM1 46 96.6
Template 2 4.2
DMSO 1.5 3.15
Phusion 0.5 1.05
50 205

Transfer 50μL of MM2 into each of 4-PCR tubes.
PCR

98°C for 3 min
36 cycles
98°C for 10s
66°C for 30s
72°C for 60s
72°C for 10 min
4°C forever


Sample g039 may have too much gDNA, samples g045 and g046 were in non-autoclaved tubes, sample g046 did not have 50ul for each tube