For in-depth product & services help, ask our
Technical Information Scientists
Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination. The transgene genotype is determined by comparing ΔCt values of each unknown sample against known homozygous and hemizygous controls, using appropriate endogenous references.
Mut= 108 bp
Wt= 120 bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
31583 | TGA CTG GTA GCT GTA AAG TCA TGG | Common | A | |||
31584 | TTC CAC ACC CAA GTA GAC ATC | Wild type Reverse | A | |||
31585 | TTG CTG GGC AAC AAT GTC | Mutant Reverse | A | |||
31586 | Fluorophore-1 | AGG TCA CAG CTC CCC TGA G | Quencher-1 | WT Probe | ||
31587 | Fluorophore-2 | AAG ATG TGT TGG AGC ACC ATT | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
31583 | 0.40 uM |
31584 | 0.40 uM |
31585 | 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 |