Reagent | Reagent Volume(µL per sample) | Final Conc. |
|
Mix A | Mix B | ||
Taq PCR Mix (2×) | 10 | 25 | 1× |
Forward Primer (10 μM) | 0.5 | 1.0 | 0.25 μM |
Reverse Primer (10 μM) | 0.5 | 1.0 | 0.25 μM |
DNATemplate* | Variable | Variable | / |
PCR Grade H2O | To 20 | To 50 | / |
Total volume per reaction | 20 | 50 |
DNA Template | Reagent Volume |
基因组DNA | 20 ng ~ 100 ng |
质粒DNA | 50 pg ~ 10 ng |
cDNA(产物原液) | 1 μL ~ 5 μL (不超过反应体积的1/10) |
Initial Denaturation | 95℃ | 2 min | ||
Denaturation | 95℃ | 25 sec | 35-40循环 | |
Annealing*1 | 60℃ | 25 sec | ||
Extension*2 | 72℃ | 1 min | ||
Final Extension | 72℃ | 7 min |