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.
>chr8:126397180-126397328 149bp AGAAGCCATGTCCCCAGA TCTCAAATGCACGCCTTC
Mutant= 152 bp
Wild Type = 149 bp
Wt Sequence: agaagccatgtccccagagctagccttactgtgtaagaccgcttcttcccaggacttctggacaactgaggcgtctgagtcccctgcagaccagctgttggtatccttggcacatggctctgagccctccagaaggcgtgcatttgaga
Mut Sequence: CACTGTGTATCCTGGAAGTTTATTTATTACTAACTCGCAGATGTCTTTGccGTGGTCGCCATTGCTGTTGCCCCGGGCGACGCCGCCAGCTCGCTCCGCCAACCGGAAGCGGGAgcgagggtcacgtgaccggccgcgccctcgcgagacttgacttccggc
A 2611 bp deletion beginning at Chromosome 8 position 125,669,882 bp and ending after 125,672,492 bp (GRCm38/mm10).
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
67619 | GCC GGA AGT CAA GTC TCG | Mutant Reverse | A | |||
67620 | Fluorophore-1 | AGA TGT CTT TGC CGT GGT CGC CAT | Quencher-1 | MUT Probe | ||
67793 | CCT GGA AGT TTA TTT ATT ACT AAC TCG | Mutant Forward | A | |||
67794 | AGA AGC CAT GTC CCC AGA | Wild type Forward | A | |||
67795 | TCT CAA ATG CAC GCC TTC | Wild type Reverse | A | |||
67796 | Fluorophore-2 | ACA ACT GAG GCG TCT GAG TCC | Quencher-2 | WT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
67619 | 0.40 uM |
67793 | 0.40 uM |
67794 | 0.40 uM |
67795 | 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 |