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Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.
Mutant= 79 bp
Wild Type = 83 bp
>chr10:75326318+75326400 83bp TCGACAGATACATCGCCATC TTTGTGCCCACAGATCTAGC
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
59285 | TCG ACA GAT ACA TCG CCA TC | Common | A | |||
59286 | TTT GTG CCC ACA GAT CTA GC | Wild type Reverse | A | |||
59287 | Fluorophore-1 | TCC ACT CCG GTG AGC CAG | Quencher-1 | WT Probe | ||
59288 | Fluorophore-2 | TCT ACC GGG TAG GGG AGG C | Quencher-2 | MUT Probe | ||
oIMR5316 | CTA AAG CGC ATG CTC CAG AC | Mutant Reverse | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
59285 | 0.40 uM |
59286 | 0.40 uM |
oIMR5316 | 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 |