05/18/2010: Oops, change of plans
- When I was just about to quantify the double terminator with the purified luciferase(pur.luc.), I realized that the band size of the second one was not the expected one (~1600 bp). In order to discard the fact that the band observed was that of the plasmid, but not the luciferase per se, I ran a gel of the pur.luc. and of the strain with the plasmid and luciferase. The results showed the plasmid and the luciferase, but the pur.luc. wasn't there.
- In order to make sure that the next steps of the week plan are achieved successfully (ligation, transformation, induced mutation, etc), I decided to repeat the band purification.
- I made a double restriction using ECOR1 and SPEI, and incubated ON at 37°C. The reaction (40μL) was done by duplicate using as DNA input the previously used strains of Arturo Zepeda. It was stored as Luciferasa restriccion ECO RI y SPE I para purificación (1-B).
-H2O ------> 6μl
-BSA ------> 1μl
-Buffer ---> 4μl
-ECO RI --> 2μl
-SPEI -----> 2μl
-DNA ------> 25μl