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Primer | 5' Label | Sequence 5' → 3' | 3' Label | Primer Type | Reaction | Note |
---|---|---|---|---|---|---|
oIMR1110 | CAA GAC AAT ATT CCT GGT GCC | Reverse | ||||
oIMR1163 | CAA TTT TGA GGA ATC TAA GGC C | Forward |
Component | Final Concentration |
---|---|
ddH2O | |
AB PCR BufferII | 1.00 X |
MgCl2 | 2.00 mM |
dNTP | 0.20 mM |
oIMR1163 | 1.00 uM |
oIMR1110 | 1.00 uM |
DNA Loading Dye | 0.69 |
Taq DNA Polymerase | 0.03 U/ul |
DNA |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 94.0 | -- | |
2 | 94.0 | -- | |
3 | 55.0 | -- | |
4 | 72.0 | -- | repeat steps 2-4 for 35 cycles |
5 | 72.0 | -- | |
6 | 10.0 | -- | hold** |
Component | Final Concentration |
---|---|
Rest Enzyme Buffer | 1.00 |
ddH2O | |
Stu I | 0.83 U/ul |
PCR product |
Step | Temp °C | Time | Note |
---|---|---|---|
1 | 37.0 | -- | 2-6 hours |
2 | 10.0 | -- | hold |