5.2 Genotyping Salk mutants (PCR)

5.2 Genotyping Salk mutants (PCR) kdorfman Thu, 03/24/2022 - 18:15

Check extracts for DNA

  • 1% gels
  • HW Massruler

Amplify extracted DNA 24 PCR reactions per pair (12 WT + 12 Salk)

Reagent WT allele master mix uL mutant master mix uL per pair per class
H2O 243.25 243.25 486.5 ~6 mL
10X ex taq buf 35 35 70 ~145 uL
dNTP 28 28 60 720 uL
LBP 17.5 17.5 210 uL
GSP 1 17.5 ?
GSP 2 17.5 ?
Taq 5U/uL 1.75 uL 1.75 uL 3.5 uL 42 uL