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Mutant = G/G
Heterozygote = C/G
Wild type = C/C
AAACATGATTTTTTAGACAACTGTCACTACACAGGATACTTGCAAGATCAACA
TAGTACAACTAAAGTGGCTTTAAG(C/G)AACTGTGTTGGATTGgtaagtaaatataatt
Ttaaatttctagtgggaaaataaattcacaatgtaaatgtctttcctcagcttaaaataagtagataaataac
atttaagtagaaagataataattttatgttcattgtttctgcttctcttacctttatttttgctattgtaatcttg
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
18560 | GAC CCC AGT GGA AAC ATG A | Forward | A | |||
18561 | GCT GAG GAA AGA CAT TTA CAT TGT G | Reverse | A | |||
18562 | Fluorophore-1 | GTG GCT TTA AGC AAC TGT GTT GG | Quencher-1 | WT Probe | ||
18563 | Fluorophore-2 | GTG GCT TTA AGG AAC TGT GTT GG | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
18560 | 0.40 uM |
18561 | 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 |