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Mutant =A/A
Heterozygote = T/A
Wild type = T/T
>chr5:30696961-30697067 107bp TCGACTTCCATGTCTCTCCTG TCCTGCGTTAGTCAGTGTGG
SEQ
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
40489 | Fluorophore-1 | TCG ACT TCC ATG TCT CTC CTG | Quencher-1 | Forward | A | |
40490 | Fluorophore-2 | TCC TGC GTT AGT CAG TGT GG | Quencher-2 | Reverse | A | |
40491 | Fluorophore-3 | TCA TCA AGA TCT CGG TGA GTG G | Quencher-3 | WT Probe | ||
40492 | Fluorophore-4 | TCA TCA AGA ACT CGG TGA GTG G | Quencher-4 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
40489 | 0.40 uM |
40490 | 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 |