November 13, 2012
- Digests:
- In a 30 μL reaction, cut H2B with XbaI and SpeI
- In a 30 μL reaction, cut LOV with XbaI and SpeI
- In a 30 μL reaction, cut a pSB1A2 plasmid (from Rene) XbaI and SpeI
| Reagent | H2B | LOV | pSB1A2
|
| DNA | 8.0 | 8.0 | 15.0
|
| XbaI | 1.0 | 1.0 | 1.0
|
| SpeI | 1.0 | 1.0 | 1.0
|
| 10x buffer | 3.0 | 3.0 | 3.0
|
| dH2O | 10.0 | 10.0 | 10.0
|
| Total vol. | 23. 0 | 23.0 | 30.0
|
- Run entire 30 μL on a 1% gel (use the big fat-tooth well comb)
- Cut band out of gel (use UV box); H2B = ~410, LOV = ~461, pSB1A2 = ~2000
Image:VN Gel 11-13-12.png
Results of the enzyme digests are shown, with H2B on the left, LOV in the middle, and pSB1A3 on the right. The gel was loaded with 30 μL samples.
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