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.
>chr8:57487831-57487950 120bp CAGAGCGCAAGTTAGCTCCT CCTCCTTCTCAACCCCAGA
Mut = 105 bp
Wt = 120 bp
Fam = Mut
Hex = Wt
MUT Sequence (junction in uppercase):
gaacagagcgcaagttagctcctcgatcaagaactaCCttccctaggtggatttgatttgcaattaagcattcttgtttttgttttgttttgtttctccgtgtaaccctggatgccct
This mutation is a 5368 bp deletion beginning at Chromosome 8 position 57,482,549 bp and ending after 57,487,916 bp (GRCm38/mm10).
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
51839 | CAG AGC GCA AGT TAG CTC CT | Common | A | |||
51840 | CCT CCT TCT CAA CCC CAG A | Wild type Reverse | A | |||
51841 | GGG TTA CAC GGA GAA ACA AAA CA | Mutant Reverse | A | |||
51842 | Fluorophore-1 | CAG AGG AGG GGC GTC TCC | Quencher-1 | WT Probe | ||
51843 | Fluorophore-2 | ACC TTC CCT AGG TGG ATT TGA TTT G | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
51839 | 0.40 uM |
51840 | 0.40 uM |
51841 | 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 |