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Mutant = TG
Heterozygote = CA/TG
Wild type = CA
>chr5:138176867+138176937 71bp CTTCCGGCTCTTTCCCTCT TTCTAAAACACAGGAAATAGTCTCAC
ATACCAGCTCTCTGATGACCTCCCCTCACCACTCCCCTTCCGGCTCTTTCCCTCTGTGCAGTGGGAC(c/t)(a/g)AGGCTCAGGCCGGTGAGACTATTTCCTGTGTTTTAGAACTACTCCATGCCATCCCGAGGCTTACAT
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
53447 | CTT CCG GCT CTT TCC CTC T | Forward | A | |||
53448 | TTC TAA AAC ACA GGA AAT AGT CTC AC | Reverse | A | |||
53449 | Fluorophore-1 | CAG TGG GAC CAA GGC TCA G | Quencher-1 | WT Probe | ||
53450 | Fluorophore-2 | AGT GGG ACT GAG GCT CAG G | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTPS-kapa | 0.26 mM |
53447 | 0.50 uM |
53448 | 0.50 uM |
Glycerol | 6.50 % |
Dye | 1.00 X |
Kapa 2G HS taq polym | 0.03 U/ul |
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 |