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= 79 bp
Wild Type = 71 bp
chr13:114458716-114458786 71bp ACCGCAAACCTCGGAGAC GATTCAATGGACGTCAGAAGC
Wt Sequence: cctcccacagccccacacactgggagaccgcccaccGCAAAcctcggagacccccgtctaGAtttaaagcgcggctgcgcccggcttctGACGTCCATTGAATCGCGCGGGCGGCCGGTGGCGAGCGCGGGGCTGCGCCGGGATCGCTGCGCCCTCCGCCGCTCGCCTCTGCGACGCGCGCCGCTCGCCCGAGCCACCCGCCGCCGCGCTCCTCGCCCCGCGcctgccccaggATGGTCTGCGCCAG
Mutant Sequence: CCCACACACTGGGAGACCGCCCACCGCAAACCTCGGAGACCCCCGTCTAGcccatcgataagcttgatatcgaattctaccgggtaggggaggcgcttttcccaaggcagtctggagcatgcgctttaa
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
41272 | GAT TCA ATG GAC GTC AGA AGC | Wild type Reverse | A | |||
53881 | ACC GCA AAC CTC GGA GAC | Common | A | |||
53884 | Fluorophore-1 | CCC GTC TAG CCC ATC GAT AAG C | Quencher-1 | MUT Probe | ||
53885 | Fluorophore-2 | CCC GTC TAG ATT TAA AGC GCG G | Quencher-2 | WT Probe | ||
oIMR6592 | GAA AAG CGC CTC CCC TAC | Mutant Reverse | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
41272 | 0.40 uM |
53881 | 0.40 uM |
oIMR6592 | 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 |