User:Mar/Notebook/2007-8-6

= Optimization of PCR cycles for genotyping (2) = Goal: shorten PCR cycling while preserving detection level

GreenGoTaq - 75µL primers - 7.5µL each water - 45µL rl117b (het) DNA - 7.5µL
 * prepared 8x20µL = 150µL of mix:
 * aliquoted : 4x5µL; 4x10µL; 4x20µL, added 10µL mineral oil to 5µL and 10µL tubes and frozen all tubes on dry ice
 * started AWS10, AWS15 (4:07pm-5:33) and AWS30 (4:10pm-6:00) on sets of tubes, then frozen amplificates until next day
 * started AWS with another set of tubes o/n, then frozen in the morning
 * (next day) thawed and run gel

ramp time for PTC-200: up to 3°C/sec

actual time @ 20µL: 1°C/sec

programs used: