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Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
oIMR5265 | TTC AGA TCT GAC CTC AGA CCC | |||||
oIMR5266 | AGC TTC TTG ATC GCA CCG TTC |
Component | Final Concentration |
---|---|
ddH2O | |
AB PCR BufferII | 1.00 X |
MgCl2 | 2.00 mM |
dNTP | 0.20 mM |
oIMR5265 | 1.00 uM |
oIMR5266 | 1.00 uM |
DNA Loading Dye | 0.69 mM |
Taq DNA Polymerase | 0.03 U/ul |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 94.0 | -- | |
2 | 94.0 | -- | |
3 | 58.0 | -- | |
4 | 72.0 | -- | repeat steps 2-4 for 35 cycles |
5 | 72.0 | -- | |
6 | 10.0 | -- | hold** |
Component | Final Concentration |
---|---|
ddH2O | |
AB PCR BufferII | 5.45 X |
BsoB1 |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 37.0 | -- | Add 2.2 ul of buff/Enzyme to entire PCR product |