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Mutant = T
Heterozygote = -/T
Wild type = -
Mutation is a T insertion.
WT sequence showing the {insertion}:
ACTACCAACCAACTCCACAGAAATGGCCGGGGCCTCTGTGAAGGTGGCGGTGCGTGTCCGCCCCTT{t}CAACTCCCGGGAAATGAGCCGAGACTCCAAGTGCATCATTCAGATGTCTGGAAGCACCACCA
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
52892 | GGG GCC TCT GTG AAG GTG | Forward | A | |||
52893 | TTG GAG TCT CGG CTC ATT TC | Reverse | A | |||
52894 | Fluorophore-1 | CGC CCC TTC AAC TCC C | Quencher-1 | WT Probe | ||
52895 | Fluorophore-2 | CGC CCC TTT CAA CTC CC | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
52892 | 0.40 uM |
52893 | 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 |