Haynes Lab:Notebook/CRISPR Editing/2015/02/06

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02/06/2015

Trying Surveyor on g023 with a shorter transcript. Still probably too long but I know these primers work and it's still like half of the transcript I was using before. First, PCR with phusion:

' 1rxn 6.2
H2030.8190.96
HF 5X MasterMix1062
dNTPs0.21.24
FP P1592.515.5
RP P1702.515.5
Total46285.2
Cell type treatment rep ng/µL ul for 200ng
Luc14g023182.3582.43
Luc14g023276.4372.62
Luc14g0233119.2811.68
GAL4-EED +doxg0231122.8441.63
GAL4-EED +doxg0232102.4341.95
GAL4-EED +doxg0233101.8621.96
' 1rxn
MM146
Template2
DMSO1.5
Phusion0.5
Total50

PCR

98°C for 3 min
30 cyles
98°C for 10s
69°C for 30s
72°C for 30s
72°C for 10 mins
4°C



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