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= 120 bp
Wild Type = 122 bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
31225 | TTT AAG GTA TCT TGT TTT AAG GTT TTG | Common | A | |||
31226 | Fluorophore-1 | AAA TGA GTG GGG TGG TGT TT | Quencher-1 | MUT Probe | ||
31227 | AGA AGG ACT GTC TGT CAC TTT GC | Wild type Reverse | A | |||
31228 | TGT GCC CAG CCT GCT TAC TA | Mutant Reverse | A | |||
31229 | Fluorophore-2 | TTT CAG TTG ATA GGG GAA ACG | Quencher-2 | WT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
31225 | 0.40 uM |
31227 | 0.40 uM |
31228 | 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 |