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.
Mutant= 116 bp
Wild Type = 100 bp
>chr4:43037233-43037332 100bp CTGCAGAGCTTGTCCCAAA AGAATCCGTGCAGCCAAT
Large deletion: Mutant sequence with junction in uppercase
tctgttgagtcctcaggtagcccaattcttctgcatacgccctaggtctccagcctgcctcttctgtttctgcagagcttgtcccaaaGAtcttaggctgggatgagctgagtcttggaccatagtcttagtcctttttgtataaggtgctgggggctgggtgggtaaagtgaagctacagaggctaaactctgagaatctggattcttacactgattctgtagtttagccccat
This mutation results in a 4988 bp deletion beginning at Chromosome 4 position 43,032,325 bp and ending after 43,037,312 bp (GRCm38/mm10).
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
66394 | CTG CAG AGC TTG TCC CAA A | Common | A | |||
66395 | AGA ATC CGT GCA GCC AAT | Wild type Reverse | A | |||
66396 | Fluorophore-1 | AGA TGT CTT GGT GTG TGT GCT G | Quencher-1 | WT Probe | ||
66397 | Fluorophore-2 | AGG CTG GGA TGA GCT GAG TCT TGG | Quencher-2 | MUT Probe | ||
66398 | GCC TCT GTA GCT TCA CTT TAC CC | Mutant Reverse | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
66394 | 0.40 uM |
66395 | 0.40 uM |
66398 | 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 |