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.
>chr13:21981553+21981639 87bp CTCTCAGCATGGGCAGCTA CAGGCTTCTCCCCTGAGTCT
Mutant= 81 bp
Wild Type = 87 bp
Wt Sequence: ctctcagcatgggcagctacctgggCAAAGCGGGGTCCTCGCCGCGGTCCCCAGCACAGGGGCGCGCAGACTCAGGGGAGAAGCCTG
Mut Sequence: CTCTCAGCATGGGCAGCTACCTGGgcATCACACTTACAAGAAATAGCCCACTCTCTCTGTCACACTGCAACAGAGTCTTGA
A 2881 bp deletion beginning at Chromosome 13 position 21,981,578 bp and ending after 21,984,458 bp (GRCm38/mm10).
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
65247 | CTC TCA GCA TGG GCA GCT A | Common | A | |||
65248 | CAG GCT TCT CCC CTG AGT CT | Wild type Reverse | A | |||
65249 | TCA AGA CTC TGT TGC AGT GTG A | Mutant Reverse | A | |||
65250 | Fluorophore-1 | TGG GCA TCA CAC TTA CAA GAA ATA | Quencher-1 | MUT Probe | ||
65251 | Fluorophore-2 | TGG GCA AAG CGG GGT | Quencher-2 | WT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
65247 | 0.40 uM |
65248 | 0.40 uM |
65249 | 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 |