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Tg= 72 bp
IPC= 74 bp
Plese note this is an inverse qPCR where the wt will have two copies (hom-like); and homs will have no copies (wt-like)
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
40496 | Fluorophore-1 | AAA GGC AGC AAA AAC AAG AGC | Quencher-1 | MUT Probe | ||
oIMR1544 | CAC GTG GGC TCC AGC ATT | Internal Positive Control Forward | A | |||
oIMR3580 | TCA CCA GTC ATT TCT GCC TTT G | Internal Positive Control Reverse | A | |||
oIMR5139 | CCT AAG AGA CCC AGA GGA AGA CC | Mutant Forward | A | |||
oIMR5140 | TCT CAC CTT CTG GGC TGC TT | Mutant Reverse | A | |||
TmoIMR0105 | Fluorophore-2 | CCA ATG GTC GGG CAC TGC TCA A | Quencher-2 |
Component | Final Concentration |
---|---|
Kapa Probe Fast QPCR | 1.00 X |
ddH2O | |
oIMR1544 | 0.40 uM |
oIMR3580 | 0.40 uM |
oIMR5139 | 0.40 uM |
oIMR5140 | 0.40 uM |
Tg Probe | 0.15 uM |
IC Probe | 0.15 uM |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 95.0 | -- | |
2 | 95.0 | -- | |
3 | 60.0 | -- | repeat steps 2-3 for 40 cycles |