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Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
9875 | AAA TTG GGT TAC ACC GCT GA | Common | ||||
9877 | CCA CCT AGC CTG CCT GTA CT | Mutant Reverse | ||||
oIMR7172 | CTG TAT CCT TGG GTC AAG CTG | Wild type Reverse |
Component | Final Concentration |
---|---|
ddH2O | |
KAPA Rxn Buffer Contains 1.5 mM Mg2+ final concentration. | 1.00 mM |
dNTP | 0.20 |
9875 | 1.00 |
9877 | 1.00 |
oIMR7172 | 1.00 |
DNA Loading Dye | 0.69 |
KAPA LR HotStart Taq Initial Denature 3 Min This is a two enzyme system. One with proofreeading the other without. HotStart is provided by antibody. | 0.01 mM |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 94.0 | -- | |
2 | 94.0 | -- | |
3 | 65.0 | -- | |
4 | 72.0 | -- | repeat steps 2-4 for 35 cycles |
5 | 72.0 | -- | |
6 | 10.0 | -- | hold |