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November 9, 2012
- First, resuspend dry primer oligos (in blue-capped tubes) to 100 μM
- Then make working solutions of 10 μM
- Reactions:
- H2B-GFP plasmid + BB_H2B fwd + BB_H2B rev
- LOV plasmid + BB_LOV fwd + BB_LOV rev
| Reagents |
H2B |
LOV
|
| Plasmid DNA |
0.5 μL |
0.5
|
| primer 1 (10 μM) |
1.0 |
1.0
|
| primer 2 (10 μM) |
1.0 |
1.0
|
| 2x GoTaq mix |
12.5 |
12.5
|
| dH2O |
10.5 |
10.5
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PCR program:
- 95°C, 3 min.
- [95°C, 30 sec; 57°C, 30 sec.; 72°C, 30 sec.] x35 cycles
- 72°C, 3 min.
- 4°C ∞
- Results of the PCR product digests are shown, with H2B on the left and LOV on the right.
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