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PCR Genotyping for Gene Targeting F1 Mice

PCR Conditions for Genotyping

You will receive primer sequences for the gene targeting mice. The following conditions are used for PCR genotyping for all our gene targeting mice. We'll email you if special conditions are used for your mice.

PCR Mix - Stocks
One Reaction
Ten Reactions (make for 11 reactions)
dNTP 10mM 0.25 l 2.75 l
10x PCR Buffer 1.25 l 13.75 l
Primers 10 uM 0.3 l each 3.3 l each
Taq Polymerase 0.06 l 0.66 l
H2O 9.48 l 104.28 l
Total Volume 12.02 l 124.74 l
Add DNA: 0.5 l    

PCR Cycles
1. Denaturation: 94C 3 minutes
2. Denaturation: 94C 30 seconds
3. Annealing: 55 C 30 seconds
4. Extension: 2-4 steps X 35 cycles 72 C 30 seconds
5. Extension 72C 5 minutes