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Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.
Mut= 95 bp
Wt= 108 bp
GTGGCTCAGGACACTCCCCCCacggagctgcattTTGGGGAGAAATGGTTCCACAAGAAG
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
29569 | AAT AAG CCC TTT CGG ATG CT | Forward | A | |||
37723 | CTT CTC CGC ACT GGT CCT AC | Reverse | A | |||
37724 | Fluorophore-1 | CAC GGA GCT GCA TTT TGG | Quencher-1 | WT Probe | ||
37725 | Fluorophore-2 | CCT TGG GGA GAA ATG GTT C | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
29569 | 0.40 uM |
37723 | 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 |