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Mutant = 72 bp
Wild type = 113 bp
Fam=Mut
Hex=WT
>chr11:119135923+119136035 113bp ACTTGTTCCTGAGCCCAAAC CTGGGCCTAGGAACACATACA
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
38768 | ACG GGT GTT GGG TCG TTT | Mutant Forward | A | |||
38769 | ACT TGT TCC TGA GCC CAA AC | Wild type Forward | A | |||
38770 | Fluorophore-1 | TCG GAT CCG TCG ACA AGC TT | Quencher-1 | MUT Probe | ||
38771 | Fluorophore-2 | AGC CCA GCC CTA ACC TGG AG | Quencher-2 | WT Probe | ||
oIMR1749 | CTG GGC CTA GGA ACA CAT ACA | Common | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
38768 | 0.40 uM |
38769 | 0.40 uM |
oIMR1749 | 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 |