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Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.
Mut= 72 bp
Wt= 93 bp
This assay is NOT able to be used for copy number evaluation. If this is required, it is suggested to type by qPCR.
Wt Sequence:
CTTGCCCTCCAAGTTCATCTTTCAGTTTCTTCAGGAGGTCCCCAGCATAGGGGCTGAGGAGCTCAACATAGCCAGCTCAGCCCCTCAGACACCCCCATCAGAGTTACAGCCCTTCCTGGCACCTGTGATCACCAACCAGAGGGTCACTGGCCCCCAACACTTTCAGGATGTTCGGCTGATTgaATTTGACATTACAGACTCCAATATCAGgtaaggaacccaagccctcaggcagtcaggcagggtgcccatatattcctaggaagctgatgtggtcagggaaaaggtaggcagtagttagggcttcacaagggtatgccagagccaaaggccaggcagttccgagagcttga
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
40196 | TCA CCA ACC AGA GGG TCA CT | Wild type Forward | A | Ndor1 | ||
40197 | GGG TTC CTT ACC TGA TAT TGG A | Common | A | Ndor1 | ||
40198 | TCT TGT CTT CGT TGG GAG TG | Mutant Forward | A | vector | ||
40199 | Fluorophore-1 | TTC GGC TGA TTG AAT TTG ACA TTA | Quencher-1 | WT Probe | ||
40200 | Fluorophore-2 | ATT AGC CCT TCC AAT TTG ACA TTA | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
40196 | 0.40 uM |
40197 | 0.40 uM |
40198 | 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 |