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Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.
Mutant= 106 bp
Wild Type = 108 bp
>chr13:38039891+38039998 108bp GCAAAAGCAAAGCCAACAC CCCAATCTGCTCCGTCTTCT
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
36620 | GCA AAA GCA AAG CCA ACA C | Common | A | |||
36621 | CCA GTG GCT GGC TAA ACA GT | Mutant Reverse | A | |||
36622 | CCC AAT CTG CTC CGT CTT CT | Wild type Reverse | A | |||
36623 | Fluorophore-1 | CAG TCC TGA CAT CAC TGA GCC | Quencher-1 | MUT Probe | ||
36624 | Fluorophore-2 | TGC AGA TGA GGA GAC AGA GAA CT | Quencher-2 | WT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
36620 | 0.40 uM |
36621 | 0.40 uM |
36622 | 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 |