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Mutant = T/T
Heterozygote = C/T
Wild type = C/C
CTGGGATGACCGCATGCGCACAGTCAAGTGGGTGGATGCTCCCCGAAAT
ACACCTCTGACCTTGAATACAAAA(C/T)GAAAGgtgagtatacaagggacaggaa
acacagacttcatccagcccgccatgctc
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
18552 | AGT CAA GTG GGT GGA TGC TC | Forward | A | |||
18553 | GGG CTG GAT GAA GTC TGT GT | Reverse | A | |||
18556 | Fluorophore-1 | TTG AAT ACA AAA CGA AAG GTG AGT A | Quencher-1 | WT Probe | ||
18557 | Fluorophore-2 | CTT GAA TAC AAA ATG AAA GGT GAG TAT A | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
18552 | 0.40 uM |
18553 | 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 |