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= x bp
Wild Type = y bp
Wt Sequence:
Mutant Sequence:
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
34592 | TCC AAA AGA AGG CAA GGT CA | Common | A | |||
34593 | Fluorophore-1 | CCC GAT CTG ACC TTG TGT CT | Quencher-1 | WT Probe | ||
34594 | TCC GAG ACC TCT CCA TGC | Wild type Reverse | A | |||
34595 | Fluorophore-2 | GTC TGC GTG CCC ATT CTT T | Quencher-2 | MUT Probe | ||
34596 | CAA CAA ACA ATA AAT GAA CAA ACC A | Mutant Reverse | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
34592 | 0.40 uM |
34594 | 0.40 uM |
34596 | 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 |