User:Mariana Ruiz Velasco L./Notebook/IGEM 2010/Wet lab journal/2010/08/16

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Let's work it, and work it, let's work it...
    1 and 14. Ladder.
 * As I did Endy: Colony PCR in previous days, I ran a gel to check on it, where the lanes were ordered as follows.
 * Lanes:

2. Positive control.

3. Negative control. 

4. Luciferase 1. 

5. Luciferase 4. 

6. Luciferase 5. 

7. Luciferase 6. 

8. Luciferase 7. 

9. Mut luz 1. <br/ >

10. Mut luz 2. <br/ >

11. Mut luz 5. <br/ >

12. Mut luz 6. <br/ >

13. Mut luz 8. <br/ > <br/ > <br/ > <br/ > -H2O ---> 10μl<br/ >
 * In order to check the LRE synthesis, I purified plasmid from 3 strains with the High Pure Plasmid Isolation Kit of Roche.<br/ >
 * Finally, I did a restriction for a total of 20μL as follows:<br/ >

-Buffer 4 --> 2μl<br/ >

-BSA ---> 1μl<br/ >

-DNA --> 5μl<br/ >

-ECOR1 -> 1μl<br/ >

-PST1 --> 1μl<br/ >

<br/ > <br/ >
 * Restrictions were incubated at 37°C ON(overnight) and labeled as LRE No. R-> ECO y PST Mar.


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