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IPC = 88bp
TG = 116bp
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
14926 | AGG ATG TTC CCC AAT GCG | Wild type Forward | A | |||
14927 | CCG GAA TCC TTA CCT TGG TTG | Wild type Reverse | A | |||
14928 | Fluorophore-1 | CCT ACC TGC CCA GCT GCC T | Quencher-1 | WT Probe | ||
14929 | AGG AGC GGA GAA CCG TC | Mutant Forward | A | |||
14930 | TCT TGC GAA CCT CAT CAC TC | Mutant Reverse | A | |||
14931 | Fluorophore-2 | CTG ACC GTA CAC CAA AAT TTG CCT GC | Quencher-2 | MUT Probe |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
14926 | 0.40 uM |
14927 | 0.40 uM |
14929 | 0.40 uM |
14930 | 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 |