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= 103 bp
Wt= 113 bp
Wt Sequence:
Mutant Sequence:
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
32507 | AAG GCA CTT CCC TTT GAG CTA T | Mutant Forward | A | |||
32508 | Fluorophore-1 | TGA CCA AAT TCC TTG TTC TGT G | Quencher-1 | MUT Probe | ||
32509 | TCT GGA AAG TGC TCC ACA AT | Wild type Forward | A | |||
32510 | Fluorophore-2 | TAC AGG CCC TCC AAG GTA AG | Quencher-2 | WT Probe | ||
32511 | TCT CTC TCC CAA TTC AGA AAC C | Common | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
32507 | 0.40 uM |
32509 | 0.40 uM |
32511 | 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 |