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Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
11793 | GGC ATT GCG TAC ATT CTC CTA | Forward | ||||
11794 | Fluorophore | GAC AGT TTC CTC CAA GAG AAT CC | Reverse | |||
11795 | AGA AAT CAG CAT ATT CCT TA | SEQ |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 0.48 |
MgCl2 | 0.96 |
dNTPS-kapa | 0.10 |
11793 | 0.12 |
11794 | 0.12 |
Glycerol | 2.40 |
Kapa 2G HS taq polym | 0.01 |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 94.0 | -- | |
2 | 94.0 | -- | |
3 | 65.0 | -- | -0.5 C per cycle decrease |
4 | 68.0 | -- | |
5 | -- | repeat steps 2-4 for 10 cycles | |
6 | 94.0 | -- | |
7 | 60.0 | -- | |
8 | 72.0 | -- | |
9 | -- | repeat steps 6-8 for 28 cycles | |
10 | 72.0 | -- | |
11 | 10.0 | -- | hold |