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Mutation is a 25 bp deletion
GACCCTGACCTCTGCCAACC[aggctcccttccaggtggagccacc]TGAGACCCTCAGGAGGTACT
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
17573 | GCT GTG ACC TGA TGG CAC T | Forward | A | |||
17574 | ATG GAC AAG GGA CAG CAA AG | Reverse | A | |||
17575 | Fluorophore-1 | CTC CCT TCC AGG TGG AGC C | Quencher-1 | WT Probe | ||
17576 | Fluorophore-2 | CTC TGC CAA CCT GAG ACC CTC | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
17573 | 0.40 uM |
17574 | 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 |