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= 80 bp
Wild Type = 134 bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
32968 | Fluorophore-1 | CAC TGG GGA GCA TTT ACC AG | Quencher-1 | Internal Positive Control Forward | A | |
32971 | Fluorophore-2 | GTG GTC AAG TTT CAT CTC AGC A | Quencher-2 | Internal Positive Control Reverse | A | |
32975 | TCA GAG CCT GCT TCT GTT CTC | Transgene Forward | A | Itgax | ||
32976 | CAA TCA AGG GTC CCC AAA C | Transgene Reverse | A | beta-globin | ||
32977 | Fluorophore-3 | TGC AGG AAT TGA TCC TGA GAA CTT C | Quencher-3 | Tg Probe | ||
32981 | Fluorophore-4 | CCA TGA TTC TGG GTG CAG ACA G | Quencher-4 | IC Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
32968 | 0.40 uM |
32971 | 0.40 uM |
32975 | 0.40 uM |
32976 | 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 |