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Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
oIMR1586 | AGG CAG GAA GAT CAC GTG TTC AAG TAC | |||||
oIMR1587 | CAC ACG CAC ACT CTG ACA TGC ACA GGC |
Component | Final Concentration |
---|---|
ddH2O | |
KAPA Rxn Buffer Contains 1.5 mM Mg2+ final concentration. | 1.00 X |
dNTP | 0.20 mM |
oIMR1586 | 1.00 uM |
oIMR1587 | 1.00 uM |
DNA Loading Dye | 0.69 mM |
KAPATaq DNA Polymera No proofreading function. 1 error per 40000 nucleotides. Initial denature 2 min. | 0.04 U/ul |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 94.0 | -- | |
2 | 94.0 | -- | |
3 | 69.0 | -- | |
4 | 72.0 | -- | repeat steps 2-4 for 35 cycles |
5 | 72.0 | -- | |
6 | 10.0 | -- | hold** |
Component | Final Concentration |
---|---|
ddH2O | 0.00 |
NEB Buffer 3 | 0.86 X |
BSA | 0.09 |
BsteII | |
PCR product |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 60.0 | -- | USE MJ #7 |