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.
>chr17:35204123-35204242 120bp CCTACCAACCATCCCACTTG GCAGTGAGTTCTGCTTGCTG
Mutant= 96 bp
Wild Type = 120 bp
Wt Sequence: cctaccaaccatcccacttgtccagtgcctgctccctcagggatggggacctctgatcttgatagccccccaatgtcttgtgcctcttcccagGGTACCCCAGCAAGCAGAACTcaCTGC
Mutant Sequence: aaaagcgcctcccctacccggtagaattaattcgatatcaagcttatcgataccgtctcgacctcgactaGTACCCCAGCAAGCAGAACTCACTGCTCTGGAGAGCAAGCACGGATCGTGCCTTTCTCCGACATGGCTTCTCTTTGAGCAA
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
21722 | AAA AGC GCC TCC CCT ACC | Mutant Forward | A | |||
41941 | CCT ACC AAC CAT CCC ACT TG | Wild type Forward | A | |||
42307 | GCA GTG AGT TCT GCT TGC TG | Common | A | |||
42308 | Fluorophore-1 | CTT ATC GAT ACC GTC TCG ACC T | Quencher-1 | MUT Probe | ||
42309 | Fluorophore-2 | TGC CTG CTC CCT CAG G | Quencher-2 | WT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
21722 | 0.40 uM |
41941 | 0.40 uM |
42307 | 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 |