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Mutant = T, T
Wild type = G, G
>chr11:77982518-77982630 113bp AGGAGGCAAATCCTTCACTT AGGGCCAGCACATGAGAG
TGGAGTTAAAGAGACAGTTACCTTCTGAGAATTTGCAGCCTCCAACTTCCGGCTTGCA
GGGGAGGGGACTCAGACCATGGTAGGAGGCAAATCCTTCACTTTTTTTTTTTCTTTCT
CTAGCCCACCATTGTAGAAGCCTTGCT(g/t)G(g/t)CTATGAGATGGTGCAGGT
GGCCTGTGGGGCCTCTCATGTGCTGGCCCTGTCCACAGATGGAGAGTTATTCGCCTGG
GGCAGAGGAGATGGTGGTAAGCCCACTGCCCAGCTCCAGTGTTTAGGGTCCCTCTCA
CAACTTCTCCATCATGCTTTGTTCATTCCTAT
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
45099 | AGG AGG CAA ATC CTT CAC TT | Forward | A | |||
45100 | AGG GCC AGC ACA TGA GAG | Reverse | A | |||
45101 | Fluorophore-1 | AGC CTT GCT GGG CTA TGA GAT | Quencher-1 | WT Probe | ||
45102 | Fluorophore-2 | AAG CCT TGC TTG TCT ATG AGA TG | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
45099 | 0.40 uM |
45100 | 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 |