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November 12, 2012
- The PCR products were purified with the Zymo DNA Clean & Concentrator Kit
| PCR Product |
260 |
280 |
260/280 |
ng/μL
|
| H2B |
0.052 |
0.028 |
1.865 |
51.607
|
| LOV |
0.048 |
0.026 |
1.860 |
48.095
|
Great! Next step...
- 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 |
--- |
--- |
---
|
| Total vol. |
30. 0 |
30.0 |
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
- Extract DNA from gel
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