Haynes Lab:Notebook/CRISPR Editing/2014/09/20

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09/20/2014

PCR of gRNA 20 samples

Samples
gRNA 20 + dox 1
gRNA 20 + dox 2
gRNA 20 - dox 1
gRNA 20 - dox 2
gRNA 20 Luc 14

Master mix 1

' 1 rxn 5.5 rxns
water 23.5 129.25
5x buffer 10 55
dNTPs 1 5.5
FP 2.5 13.75
RP 2.5 13.75
DMSO 1.5 8.25
41

add 41ul of MM 1 to each tube
add 4ul of each template to each tube (400ng)

' 1 rxn 5.5 rxns
H20 4.5 24.75
Phusion 0.5 2.75

add 5 ul of MM 2 to each tube

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


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