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.
>chr15:89531337+89531455 119bp CCGCAGAAACCAGAGAGGAC TGCTACTGTCCCTGCGTGT
Mutant= 110 bp
Wild Type = 119 bp
Wt Sequence: ccgcagAAACCAGAGAGGACCGGACGAAGCGTCTCTTCCGCCACTACacTGTGGGTTCCTATGACAGCCTCACTTCACACAGgtactggcaggggtgtggacacgcagggacagtagca
Mutant Sequence: CCGCAGAAACCAGAGAGGACCGGACGAAGCGTCTCTTCCGCCACTACagGCGCGCCTAGTCGACATAACTTCGTATAGCATACATTATACGAAGTTATAGCGCGAAGTTC
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
19043 | GAA CTT CGC GCT ATA ACT TCG | Mutant Reverse | A | |||
40240 | CCG CAG AAA CCA GAG AGG AC | Common | A | |||
40241 | TGC TAC TGT CCC TGC GTG T | Wild type Reverse | A | |||
40242 | Fluorophore-1 | CGC GCC TAG TCG ACA TAA CT | Quencher-1 | MUT Probe | ||
40243 | Fluorophore-2 | TGG GTT CCT ATG ACA GCC TC | Quencher-2 | WT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
19043 | 0.40 uM |
40240 | 0.40 uM |
40241 | 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 |