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Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.
Mutant= 75 bp
Wild Type = 83 bp
>chr16:35279300+35279382 83bp GAACCCAGAGGATGAAGTGG GGACGTGTTCAGATCGCAGT
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
46480 | Fluorophore-1 | CAT CGA TGC CAG GAG CAT | Quencher-1 | WT Probe | ||
46482 | GAA CCC AGA GGA TGA AGT GG | Common | A | |||
46483 | GGA CGT GTT CAG ATC GCA GT | Wild type Reverse | A | |||
46486 | CTA TGC CCT GCC TTG CTT CT | Mutant Reverse | A | |||
46487 | Fluorophore-2 | CGA GTT TCT GGC ATC AGG AG | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
46482 | 0.40 uM |
46483 | 0.40 uM |
46486 | 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 |