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Mutant = --------------------/--------------------
Heterozygote =cgtgcggagcagcgagagcc/--------------------
Wild type = cgtgcggagcagcgagagcc/cgtgcggagcagcgagagcc
Mutation is a 20bp deletion
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
19050 | CCC TCC TTC GAG TTC AAG TG | Forward | A | |||
19051 | AGG TCC CTG TTC CTC AGT CA | Reverse | A | |||
19052 | Fluorophore-1 | TGC GGA GCA GCG AGA G | Quencher-1 | WT Probe | ||
19053 | Fluorophore-2 | GAA AAG AGG CCA AAC AGA TGT GT | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
19050 | 0.40 uM |
19051 | 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 |