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= 119 bp
Wild Type = 116 bp
>chr9:66830011-66830126 116bp TGCAATAAGTAGACCAACAGTAATAAG ACTGCATGTTCAAGGCCATC
Wt Sequence: tgcaataagtagaccaacagtaataagtatttgttccttgatatttaaaatgtattttatgttggctttttgagTAgatatcactgtatagaccaggatggccttgaacatgcagt
Mutant Sequence: GCCAGGGGGATCTTGAAGTTCCTATTCCGAAGTTCCTATTCTCTAGAAAGTATAGGAACTTCAAGCTCGATACCGTCgaGTTATCACTGTATAGACCAGGATGGCCTTGAACATGCAGT
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
31786 | GCC AGG GGG ATC TTG AAG T | Mutant Reverse | A | |||
43599 | ACT GCA TGT TCA AGG CCA TC | Common | A | |||
43600 | TGC AAT AAG TAG ACC AAC AGT AAT AAG | Wild type Forward | A | |||
43601 | Fluorophore-1 | AGG AAC TTC AAG CTC GAT ACC GT | Quencher-1 | MUT Probe | ||
43602 | Fluorophore-2 | TGT ATT TTA TGT TGG CTT TTT GAG T | Quencher-2 | WT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
31786 | 0.40 uM |
43599 | 0.40 uM |
43600 | 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 |