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Mutant = 81 bp
Heterozygote = 81 bp and 105 bp
Wild type = 105 bp
>chr18:11053602+11053706 105bp TCTCTTCACCGAGGCATCC GCTAACTCTGAGCCGGTGAC
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
34487 | GCT AAC TCT GAG CCG GTG AC | Common | A | |||
35185 | ACA AAG TGG TTG ATA TCT CTA TAG TCG | Mutant Forward | A | |||
35186 | TCT CTT CAC CGA GGC ATC C | Wild type Forward | A | |||
35187 | Fluorophore-1 | CAG GGT GAG ACA GCA TTA ACA CA | Quencher-1 | WT Probe | ||
35188 | Fluorophore-2 | CAG TAG GCG GTT AAC ACA CAT TAA AT | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
34487 | 0.40 uM |
35185 | 0.40 uM |
35186 | 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 |