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Mutant = A/A
Heterozygote = G/A
Wild type = G/G
CAACACAACCGCCAttgctGAAGCCTGGGCCCGTCTGGACCACAAGTTTGACTTGATGTATGC
CAAGAGGGCTTTTGTACACTG(g/a)TATGTGGGTGAGGGAATGGAGGAGGGGGAGTTCTCTG
AGGCTCGAGAGGATATGGCTGCCCTGGAAAAAGATTATGA
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
24785 | ACA ACC GCC ATT GCT GAA G | Forward | A | |||
24786 | CCA TAT CCT CTC GAG CCT CA | Reverse | A | |||
24787 | Fluorophore-1 | TTT TGT ACA CTG GTA TGT GGG TGA G | Quencher-1 | WT Probe | ||
24788 | Fluorophore-2 | TTT TGT ACA CTG ATA TGT GGG TGA G | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
24785 | 0.40 uM |
24786 | 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 |