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Taqman qPCR protocols are run on a real time PCR instrument. Use an appropriate instrument specific Fluorophore/Quencher combination.
Mutant= 86 bp
Wild Type = 71 bp
>chrX:70962450+70962520 71bp TGGAGTCCACAAAATCTACCC CAGCTGGCACCTGAGGAC
X-linked
897 bp deletion
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
46678 | TGG AGT CCA CAA AAT CTA CCC | Common | A | |||
46679 | CAG CTG GCA CCT GAG GAC | Wild type Reverse | A | |||
46680 | Fluorophore-1 | AGT ACG GCA GTG TCT GAC TCC | Quencher-1 | WT Probe | ||
46685 | TAG CCA TCC AAA GCC TAC CC | Mutant Reverse | A | |||
46686 | Fluorophore-2 | TAG ATA GAC ACT GGG ACA AGC TTG | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
46678 | 0.40 uM |
46679 | 0.40 uM |
46685 | 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 |