Oct 12, 2006· Reactions (25 μl total) were composed of 1× PCR buffer, 4 mM MgCl 2, 2.5 mM dNTPs, 4 μM each primer, 5 U Taq polymerase (all from Roche), and 2 μl of cDNA. PCR conditions were 10 min at 95°C; 40 cycles of 30 sec at 94°C, 30 sec at 55°C, and 90 sec at 72°C; followed by 10 min at 72°C.