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= 111 bp
Wild Type = 77 bp
>chrX:148364217+148364293 77bp GCAGACAAGGATGTCTTTAATGC TGCAGGCAAATTTCTATTCC
Wt Sequence: tgctgacttagtcaccaagttttaggcagacaaggatgtctttaatgcctttgcagtgcaagcTTaatatattattaggctaggaatagaaatttgcctgcaaagcagattggtaaccctgggtttgggaggcaccatgtgcttgagtgccaccccccactccacccccagtcagtaattgttc
Mutant Sequence:CTGCTGACTTAGTCACCAAGTTTTAGGCAGACAAGGATGTCTTTAATGCCTTTgcAGTGCggatccataacttcgtatagcatacattatacgaagttaTAATATATTATTAGGCTAGGAATAGAAATTTGCCTGCAAAGCAGATTGGTAACCCTGGGTTTGGGAGGCACCATGTGCTT
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
39118 | GCA GAC AAG GAT GTC TTT AAT GC | Forward | A | |||
39119 | TGC AGG CAA ATT TCT ATT CC | Reverse | A | |||
39120 | Fluorophore-1 | CAG TGC GGA TCC ATA ACT TCG TAT | Quencher-1 | MUT Probe | ||
39121 | Fluorophore-2 | TGC AGT GCA AGC TTA ATA TAT TAT TAG GC | Quencher-2 | WT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
39118 | 0.40 uM |
39119 | 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 |