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.
>chr3:89651915+89652004 90bp TTCCCTTGAAATGCCAACAC TCAGAGGGTCCCAGGAGAGA
Mut= 115 bp
Wt= 90 bp
Fam=Mut
Hex=Wt
Wt Sequence:
ttagagactcaaagtcttataccattcccttgaaatgccaacactaagcctggggcacaccaagtcttgtgttttacctcctaactgtgtgttttctctcctgggaccctctgacgtcctc
Mutant Sequence:
taccattcccttgaaatgccaacactaagcctATAACTTCGTATAGCATACATTATACGAAGTTATATACTCTGTATCCTGTTTTCAGTTCATAGCCCATCTGTAGGGCGCAGTAGTCCAGGGTTTCCTTGATG
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
53820 | TGG ACT ACT GCG CCC TAC AG | Mutant Reverse | A | |||
53822 | Fluorophore-1 | ATC CTG TTT TCA GTT CAT AGC CCA | Quencher-1 | MUT Probe | ||
54406 | TTC CCT TGA AAT GCC AAC AC | Common | A | |||
54407 | TCA GAG GGT CCC AGG AGA GA | Wild type Reverse | A | |||
54408 | Fluorophore-2 | AAG CCT GGG GCA CAC CA | Quencher-2 | WT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
53820 | 0.40 uM |
54406 | 0.40 uM |
54407 | 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 |