User:Mariana Ruiz Velasco L./Notebook/IGEM 2010/Wet lab journal/2010/05/04

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Purifying luciferase

  • As we decided to obtain the luciferase either by PCR or by band purification, we decided to proceed with the second one. To obtain the luciferase from the plasmid (that has already been purified), I realized a double restriction using ECO R1 and SPE 1 with the following reaction for a total of 40μl:

-H2O ------> 6μl
-BSA ------> 1μl
-Buffer ----> 4μl
-ECO RI --> 2μl
-SPE I ----> 2μl
-DNA ------> 25μl

  • The reaction was incubated at 37°C for 6.5 hrs. and afterwards, I did a 2% agarose gel electrophoresis for 1 hr at 85V.

  • Oops, I got confused and I didn't add the ladder, so there was no way to prove that the observed band was the desired one.