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= 182 bp
Wt= 181 bp
Wt Sequence:
Mutant Sequence:
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
34189 | TAG GCT GAG ACA TCA AGC ACA C | Wild type Forward | A | |||
34190 | CTG TGG TAT AAA CTG ACA AAG G | Wild type Reverse | A | |||
34192 | ACA GTG GGA TAT GCA CTC CA | Mutant Reverse | A | |||
34193 | Fluorophore-1 | CTT TGA GAC TGA GTT CCA TTC ATA GT | Quencher-1 | WT Probe | ||
34194 | Fluorophore-2 | CTG TAA CAC TTC AAG TCA CCA CTG | Quencher-2 | MUT Probe | ||
36002 | ACA AAT TAA CTA TTG AAC TAA GAC TTT GC | Mutant Forward | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
34189 | 0.40 uM |
34190 | 0.40 uM |
34192 | 0.40 uM |
36002 | 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 |