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Mutant = G/G and C/C
Wild type = A/A and T/T
X-linked mutation
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
14324 | GCA TAC TTT GAA AAG GTG GGA G | Forward | A | |||
14325 | TGG AGC TTT GGG AGA TTT GG | Reverse | A | |||
14326 | Fluorophore-1 | TTT TGA CTT CAC GGT AAC TGG | Quencher-1 | WT Probe | ||
14327 | Fluorophore-2 | TTT TGA CTT CGC GGT AAC CGG | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
14324 | 0.40 uM |
14325 | 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 |