Table 2 Prescribed PCR amplification program
From: The source of SYBR green master mix determines outcome of nucleic acid amplification reactions
Mix | Steps | Temperature °C | Duration | Cycles |
|---|---|---|---|---|
ABI | UDG activation | 50 | 2 min | Hold |
Activation | 95 | 2 min | Hold | |
Denature | 95 | 15 s | 40 | |
Anneal/extend | 60 | 60 s | ||
Melt curve analysis | 95 | 10 s | ||
60 | 60 s | |||
97 | 5 s | |||
BioRad | Activation | 95 | 3 min | Hold |
Denature | 95 | 15 s | 40 | |
Anneal/extend | 60 | 45 s | ||
Melt curve analysis | 95 | 10 s | ||
55 | 60 s | |||
95 | 15 s | |||
Roche | Activation | 95 | 10 min | Hold |
Denature | 95 | 10 s | 40 | |
Anneal | 60 | 10 s | ||
Extend | 72 | 10 s | ||
Melt curve analysis | 95 | 10 s | ||
65 | 60 s | |||
95 | 15 s | |||
General lab PCR program | Activation | 95 | 10 min | Hold |
Denature | 95 | 15 s | 45 | |
Anneal/extend | 60 | 60 s | ||
Melt curve analysis | 95 | 10 s | ||
55 | 60 s | |||
97 | 5 s |