PCR For Genotyping Transgenic Mice

10x pcr buffer     2.5 ul
20mM dNTP mix 0.25
50mM MgCl2   1
OL f 10um          1.25
OL r 10um          1.25
d2H2O      17.9
TAQ                  0.1

DNA (tail)           0.75

total volume per sample   25

1. Make master mix
2. Aliquot 24.2ul per tube
3. Add 0.8ul genomic tail DNA
4. Add 0.8ul controls to correct tubes

PCR machine protocol:

94C 3sec
(94/30sec> 60C/30sec > 72C/30sec) x35 cycles
72C/5min
4C/hold