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Mutant = G/G
Heterozygote = A/G
Wild type = A/A
AGCTAAAGAGGTGCTGGGTGCTTAGGTGATGGTGTCCCTGAACTCGTGCCCCAAAGAGCTCCTGATTTTGGCTCTGGTCCTCACAGAGCACTGATGACACTGAAGAAGTACCACTCTGGGCCAGCATCCTCCCTCATAGACATCCTGTTGGGCAGCTCCACCCCAAGTCCTGCCATGGAAATACGTGAGGATTTTTAATTTTCTCA(a/g)GTATGTGTGTGTGTGTGTGTGTGTGTGTGTGTGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGGGAGAGAGAGAGAGAGATGGAGGGGGAGGGAGGGAGAGAGAGAGATGGAGGGGGAGGGAGGGAGAGAGAGAGAGAGAATGAATATGTATCCACATGAGT
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
41462 | GCC ATG GAA ATA CGT GAG GA | Forward | A | |||
41464 | Fluorophore-1 | TTT TAA TTT TCT CAA GTA TGT GTG TGT GT | Quencher-1 | WT Probe | ||
41465 | Fluorophore-2 | TTT AAT TTT CTC AGG TAT GTG TGT GTG | Quencher-2 | MUT Probe | ||
42509 | CCT CCC TCC CCC TCC ATC T | Reverse | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
41462 | 0.40 uM |
42509 | 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 |