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Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.
Mutant= T, G
Wild Type = C, T
Y155Y,L156R
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
42170 | CAC ACA CTA ACC ATA TGA CAA GC | Forward | A | |||
42171 | ATC TTG GGC AGA AGG ACT GC | Reverse | A | |||
42172 | Fluorophore-1 | TTT TGC AGA CTA CCT TGT AGC TGT G | Quencher-1 | WT Probe | ||
42173 | Fluorophore-2 | TTT GCA GAC TAT CGT GTA GCT GTG | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
42170 | 0.40 uM |
42171 | 0.40 uM |
Wt Probe | 0.15 uM |
Mutant Probe | 0.15 uM |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 95.0 | -- | |
2 | 95.0 | -- | |
3 | 60.0 | -- | |
4 | -- | repeat steps 2-3 for 40 cycles | |
5 | 40.0 | -- | Forever |