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.
Mutant= 122 bp
Wild Type = 114 bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
31322 | TCC AAT ATT GAG GAT AGA AAC AA | Mutant Forward | A | |||
31323 | Fluorophore-1 | CCT TGT GTA TAC TAA GTC AAC CAA TTT | Quencher-1 | MUT Probe | ||
31324 | GTC CTG TTC CTC CAA TCT CG | Wild type Forward | A | |||
31325 | Fluorophore-2 | CTG AGG GAG GGG ATG GAG | Quencher-2 | WT Probe | ||
31326 | AGA AAT GTG AGG CAA AGC TCA | Common | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
31322 | 0.40 uM |
31324 | 0.40 uM |
31326 | 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 |