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Mutant =C/C
Heterozygote = T/C
Wild type = T/T
AGAGCCTGAGGAACGACATTGGTGCCACTg[T/C]cCATGAGCTTTCCCGAGACAAGGAAAAG
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
27129 | ACC TAT CTG GAT AAG CGT AGC A | Forward | A | |||
27130 | GAC TCT CAC CTG TGT TCT TTT CC | Reverse | A | |||
27131 | Fluorophore-1 | TGC CAC TGT CCA TGA GCT | Quencher-1 | WT Probe | A | |
27132 | Fluorophore-2 | TGC CAC TGC CCA TGA GCT | Quencher-2 | MUT Probe | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
27129 | 0.40 uM |
27130 | 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 |