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Mut = C
WT= T
CCTGGCCTTCCGAGTCTt/c
CTGTTGCTGGCTTACTGTCGTCTCGAGATTTCTTGGGTGGCCTGGCCTTCCGAGTCTtCCACTGCACACAGTACATTAGGCATGGATCTAAGCCCATGTACACA
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
22621 | Fluorophore-1 | TCC GAG TCT TCC ACT GCA C | Quencher-1 | WT Probe | ||
22622 | Fluorophore-2 | TCC GAG TCT CCC ACT GCA C | Quencher-2 | MUT Probe | ||
39414 | CTG TTG CTG GCT TAC TGT CG | Forward | A | |||
39415 | TGT GTA CAT GGG CTT AGA TCC | Reverse | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
39414 | 0.40 uM |
39415 | 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 |