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Mutant = C
Heterozygote = T/C
Wild type = T
GACACACCCGCCTACCTGTATGAGGGCCGCTGGACCCCACCTTGTTGCCTGCGCGCGCTGCGCGAGACTGCGCGCTACGTGGTGGGCGTGCTGGAGGCGGCGGGCGTGCGCTACTGGTTGGAGGGCGGCTCGCTGCTGGGTGCAGCTCGCCACGGCGACA(T/C)CATCCCTTGGgACTACGACGTAGATCTGGGCATCTACCTGGAGGACGTGGGCAACTGCGAGCAGTTGCGGGGTGCCGAAGCTGGCTCGGTAGTGGATGAACGCGGCTTTGTGTGGGAGAAGGCGGTGGAGGGCGACTTCTTCCGA
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
44729 | GTG CGC TAC TGG TTG GAG | Forward | A | |||
44730 | TCC AGG TAG ATG CCC AGA | Reverse | A | |||
44731 | Fluorophore-1 | CGG CGA CAT CAT CCC TTG G | Quencher-1 | WT Probe | ||
44732 | Fluorophore-2 | CGG CGA CAC CAT CCC TTG | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
44729 | 0.40 uM |
44730 | 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 |