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Mutant = T
Heterozygote =C/T
Wild type = C
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
15616 | Fluorophore-1 | CCA CGT GTT CTG GTT TAA CCA | Quencher-1 | MUT Probe | ||
18219 | TAG GTT GGT TTG CCT CCA TC | Forward | A | |||
18220 | CCA TCT CCT GCT TCA TGT CA | Reverse | A | |||
18221 | Fluorophore-2 | CAC GTG TTC CGG TTT AAC C | Quencher-2 | WT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
18219 | 0.40 uM |
18220 | 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 |