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Mutant = A/A
Heterozygote = G/A
Wild type = G/G
aatatctaataaaataaaattttaaaaaattaaaactgaaagagagagacaggaggccctttctcttctgca[g/a]GCATCTAAAGA
ATATGATGAGGATTCCCTCATTCCCAGCAGCCCGGCCACAGAGACCTCAGACAACATTAGCCCTGTGGCTA
GCCCGGT
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
17262 | TAG GGG ACT TTC GGG ATA GC | Forward | A | |||
17263 | AGC CAC AGG GCT AAT GTT GT | Reverse | A | |||
17267 | Fluorophore-1 | TCT CTT CTG CAG GCA TCT AAA GA | Quencher-1 | WT Probe | ||
17268 | Fluorophore-2 | TTC TCT TCT GCA AGC ATC TAA AGA A | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
17262 | 0.40 uM |
17263 | 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 |