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.
>chr4:40262264-40262414 151bp GAAATCCAGCTTGCCTTCAC GACACAAATGCCTGACTCTCC
Mut = 160 bp
Wt = 151 bp
Fam = Mut
Hex = Wt
MUT Sequence (junction in uppercase):
taaactatttaaagaatatgtaggctattatatggtatctaagagcctactgtgtgctggtacttccccaaaggTTgaacagttagttacgggcccagcttcagtggctgtagtctctcctcccccata
This mutation is a 3749 bp deletion beginning at Chromosome 4 position 40,259,541 bp and ending after 40,263,289 bp (GRCm38/mm10).
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
50088 | GAA ATC CAG CTT GCC TTC AC | Wild type Forward | A | |||
50089 | GAC ACA AAT GCC TGA CTC TCC | Wild type Reverse | A | |||
50090 | TTG TTT GCT GCT GAG CTT ATG | Mutant Forward | A | |||
50091 | GAG AGA CTA CAG CCA CTG AAG C | Mutant Reverse | A | |||
50092 | Fluorophore-1 | TGG AGC CCA TGG TTC GCT A | Quencher-1 | WT Probe | ||
50093 | Fluorophore-2 | TGT GTG CTG GTA CTT CCC CAA AG | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
50088 | 0.40 uM |
50089 | 0.40 uM |
50090 | 0.40 uM |
50091 | 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 |