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Mutant = ------------------------
Heterozygote = agggaccactattacgtaatcctg/------------------------
Wild type = agggaccactattacgtaatcctg
TCAGAAGAGTATAATAGCCATCAGGTTTTATGCGATGGAACACCTGagggaccactattacgtaatcctg/------------------------GAAACCATGACAAAGCCAAAACCCCCAGGCTCCCATCTTCAGCAGATGTGGAATTTTGTCTGAGTTTGACCCAGTATGAATCTGGATCAATGGATAGAACTGCCAATTTCAGC
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
18428 | TCA GAA GAG TAT AAT AGC CAT CAG G | Forward | ||||
18429 | GCT GAA ATT GGC AGT TCT ATC C | Reverse | ||||
18430 | Fluorophore-1 | AGG GAC CAC TAT TAC GTA ATC CTG | Quencher-1 | WT Probe | ||
18431 | Fluorophore-2 | GGA ACA CCT GGA AAC CAT GA | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 0.42 |
ddH2O | |
18428 | 0.17 |
18429 | 0.17 |
18430 | 0.08 |
18431 | 0.08 |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 95.0 | -- | |
2 | 92.0 | -- | |
3 | 60.0 | -- | |
4 | -- | repeat steps 2-3 for 40 cycles | |
5 | 4.0 | -- | Forever |