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Mutant = 107 bp
Heterozygote = 94 bp and 107 bp
Wild type = 94 bp
>chr2:26502666-26502759 94bp TGCCCTTTCCTTAAAAGTGG CCTCCCACGCCATCTTAAA
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
49117 | CCT CCC ACG CCA TCT TAA A | Wild type Reverse | A | Wt R | ||
49118 | GCT GAG CTC GAC TTC GAA TAA C | Mutant Reverse | A | Mut R | ||
49119 | Fluorophore-1 | CGC TCG AGC CCT TAA TTA ACC G | Quencher-1 | MUT Probe | ||
49120 | Fluorophore-2 | CGA GCT CAG CCC TAC ATC GAG | Quencher-2 | WT Probe | ||
oIMR6748 | TGC CCT TTC CTT AAA AGT GG | Common | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
49117 | 0.40 uM |
49118 | 0.40 uM |
oIMR6748 | 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 |