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...


  • As I did Endy: Colony PCR in previous days, I ran a gel to check on it, where the lanes were ordered as follows.


ColonyPCRMl.JPG

  • Lanes:


1 and 14. Ladder.

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.

10. Mut luz 2.

11. Mut luz 5.

12. Mut luz 6.

13. Mut luz 8.

  • In order to check the LRE synthesis, I purified plasmid from 3 strains with the High Pure Plasmid Isolation Kit of Roche.


  • Finally, I did a restriction for a total of 20μL as follows:


-H2O -------> 10μl

-Buffer 4 --> 2μl

-BSA -------> 1μl

-DNA ------> 5μl

-ECOR1 -----> 1μl

-PST1 ------> 1μl


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