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Mut = 122 bp
WT= 144 bp
SEQ
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
33822 | TGC ACC TAA AGA CTT TGT TTG G | Mutant Forward | A | |||
33823 | ATG AGA GGA AGG TCA TCT TGG A | Wild type Forward | A | |||
33824 | Fluorophore-1 | TTG ATT TGT TTC TTT TGC ACA TC | Quencher-1 | MUT Probe | ||
33825 | Fluorophore-2 | CCC GTC AGC TGC CAG TTC | Quencher-2 | WT Probe | ||
33826 | TTC CAG GAC AGC CAG AGC TA | Common | A |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
33822 | 0.40 uM |
33823 | 0.40 uM |
33826 | 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 |