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>chr4:155070675+155070771 97bp CCACTCTGCACCCTATGCT TGTTGCACCACTCGGTGA
Mutant = C, T, G
Wild type = T, C, C
CGTGTGCAGAACCCCACTCTGCACCCTATGCTTAGAGGAACGAAGACACTCCACGGCCACACCTTG(t/c)GG(c/t)(c/g)ATCTCTTCTGCTGGGAGTGCATCACCGAGTGGTGCAACACCAAGgtactcgcgctcccgggcctg
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
53879 | CCA CTC TGC ACC CTA TGC T | Forward | A | |||
53880 | TGT TGC ACC ACT CGG TGA | Reverse | A | |||
53882 | Fluorophore-1 | ACC TTG TGG CCA TCT CTT CTG | Quencher-1 | WT Probe | ||
53883 | Fluorophore-2 | ACC TTG CGG TGA TCT CTT CTG | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
53879 | 0.40 uM |
53880 | 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 |