User:Mariana Ruiz Velasco L./Notebook/IGEM 2010/Wet lab journal/2010/08/03

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Wowowoworking hard


  • Today, I repeated the restrictions done the other day with the purified PCR product of the mutated luciferase.


1. Green luciferase with ECO RI and PST I stored as luc wt R-> ECO y PST Mar and the date.

-H2O --------> 13μl
-Buffer 3 --> 3μl
-BSA -------> 1μl
-DNA -------> 10μl
-ECO I -----> 1.5μl
-PST I ------> 1.5μl

2. Mutated-luciferase with ECO RI and PST I stored as mut-luz R-> ECO y PST Mar and the date.

-H2O --------> 13μl
-Buffer 3 --> 3μl
-BSA --------> 1μl
-DNA --------> 10μl
-ECO RI ----> 1.5μl
-PST I -------> 1.5μl

  • The gel revealed that:



  • The lanes were as follows:


1. Ladder.

2. Mutated luciferase 3/50.

3. Mutated luciferase (purified product).

4. Restriction of mutated luciferase.

5. Restriction of luciferase.

6. Restriction of pBSKII (+).


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