Virginie Orgogozo
PCR from genomic DNA
(Expand High Fidelity PCR Roche #1 732 641)
in 25uL:
2.5uL buffer with MgCl2
2.5uL dNTP 2mM each (aliquots of 50 uL, 10 uL dATP 100mM, 10 uL dGTP
100mM, 10 uL dTTP 100mM, 10 uL dCTP 100mM in 460 uL H2O milliQ)
16.75uL milliQ H2O
1uL genomic DNA
1uL primer #1 10mM (keep also a 100uM dilution stock at –20∞C: if 25.19
nMoles of lyophilized oligonucleotides : add 251.9uL of H2O milliQ)
1uL primer #2 10mM
0.25uL Taq polymerase