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This assay is NOT capable of distinguishing heterozygotes from homozygotes. If this is required, it is recommended that the Generic Neo qPCR assay be run in addition to this assay.
Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
9304 | CAA AGA CTC GGG CAT TCA TC | Wild type Forward | A | |||
9305 | GGG CAT TTT CCA AAT CAT CA | Common | A | |||
oIMR0092 | AAT CCA TCT TGT TCA ATG GCC GAT C | A |
Component | Final Concentration |
---|---|
ddH2O | |
Kapa 2G HS buffer | 1.30 X |
MgCl2 | 2.60 mM |
dNTP KAPA | 0.26 mM |
9304 | 0.50 uM |
9305 | 0.50 uM |
oIMR0092 | 0.50 uM |
Glycerol | 6.50 % |
Dye | 1.00 X |
Kapa 2G HS taq polym | 0.03 U/ul |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 94.0 | -- | |
2 | 94.0 | -- | |
3 | 65.0 | -- | -0.5 C per cycle decrease |
4 | 68.0 | -- | |
5 | -- | repeat steps 2-4 for 10 cycles (Touchdown) | |
6 | 94.0 | -- | |
7 | 60.0 | -- | |
8 | 72.0 | -- | |
9 | -- | repeat steps 6-8 for 28 cycles | |
10 | 72.0 | -- | |
11 | 10.0 | -- | hold |