Thermocycle Conditions 5-21-13
2015-01-13azim58 - Thermocycle Conditions 5-21-13
Thermocycle Conditions 5-21-13
has been associated with hs tdwn dyn (hot start touchdown dynazyme)
protocol
95 C 30 s, 95 C hold, 95 C 2 min, 95 C hold, 95 C 2 min,
95 C 30 s, 65 C 30 s, 72 C 120 s,
95 C 30 s, 64 C 30 s, 72 C 120 s,
95 C 30 s, 63 C 30 s, 72 C 120 s,
95 C 30 s, 62 C 30 s, 72 C 120 s,
95 C 30 s, 61 C 30 s, 72 C 120 s,
95 C 30 s, 60 C 30 s, 72 C 120 s,
95 C 30 s, 59 C 30 s, 72 C 120 s,
95 C 30 s, 58 C 30 s, 72 C 120 s,
95 C 30 s, 57 C 30 s, 72 C 120 s,
95 C 30 s, 56 C 30 s, 72 C 120 s,
(95 C 30 s, 55 C 30 s, 72 C 120 s)X40, 72 C 5 m, 4 C hold
1st part to warm up thermocycler
2nd part to denature
3rd part to add polymerase and denature further
10 decreasing annealing temperature cycles for touchdown
amplification for 40 cycles