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Mutant =T/T
Heterozygote = G/T
Wild type = G/G
GAACGACTTCTTCCACTCCTTCCTCATC(g/t)TCTTCCGAGTGCTGTGTGGGGAGTG
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
33247 | TGG CAC ATG AAC GAC TTC T | Forward | A | |||
33248 | TCT CGA TCC ACT CCC CAC A | Reverse | A | |||
33249 | Fluorophore-1 | CCT CAT CGT CTT CCG AGT GC | Quencher-1 | WT Probe | ||
33250 | Fluorophore-2 | TTC CTC ATC TTC TTC CGA GTG C | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
33247 | 0.40 uM |
33248 | 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 |