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.
>chr1:162850466-162850572 107bp CCCAGGACACAATCAGTAAATC AGAACCTTGGGGGTAGTTCA
Mutant= 85 bp
Wild Type = 107 bp
Wt Sequence: cccaggacacaatcagtaaatcagaacttaattcttaTttgcatatgtgcattcaatagaaacaaaaaaaatagattatttgaaccttgaactacccccaaggttct
Mut Sequence: cccaggacacaatcagtaaatcagaacttaattcttaTAcctggttagtcccaggatgaactcttgtggattgtccgttaaacag
A 526 bp deletion beginning at Chromosome 1 position 162,850,009 bp and ending after 162,850,534 bp (GRCm38/mm10).
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
65252 | CCC AGG ACA CAA TCA GTA AAT C | Common | A | |||
65253 | AGA ACC TTG GGG GTA GTT CA | Wild type Reverse | A | |||
65254 | CTG TTT AAC GGA CAA TCC ACA A | Mutant Reverse | A | |||
65255 | Fluorophore-1 | TGC ATA TGT GCA TTC AAT AGA AAC | Quencher-1 | WT Probe | ||
65256 | Fluorophore-2 | TTC TTA TAC CTG GTT AGT CCC AGG A | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
65252 | 0.40 uM |
65253 | 0.40 uM |
65254 | 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 |