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Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
oIMR0033 | GCC ATC CCT TCA CGT TAG | Wild type | ||||
oIMR0034 | TTC CAC TTT GGC ATA AGG C | Common | ||||
oIMR0758 | TTC TGA GAA AGA CAG AAG TTA | Mutant |
Component | Final Concentration |
---|---|
ddH2O | |
AB PCR BufferII | 1.00 X |
MgCl2 | 2.50 mM |
dNTP | 0.20 mM |
oIMR0033 | 1.00 uM |
oIMR0034 | 1.00 uM |
oIMR0758 | 1.00 uM |
DNA Loading Dye | 0.69 mM |
Taq DNA Polymerase + | 0.02 U/ul |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 94.0 | -- | |
2 | 94.0 | -- | |
3 | 55.0 | -- | |
4 | 72.0 | -- | Go to step 2-4, 35 times |
5 | 72.0 | -- | |
6 | 10.0 | -- |