Working harder than ever with luciferases...
- Today, I inoculated luciferases from photinus pyralis (with mutated RBS) of strains 3, 5, 8 and its mutated version (strains 6,8,9) in 4ml of LB with 4μl of ampicilin and leave them growing at 37°C for 6 hours.
- After extracting plasmid with the High Pure Plasmid Isolation Kit from Roche®, I did a restriction for a total of 30μL as follows to check if the luciferase was ligated in the pBSIIK(+/-):
-H2O -------> 13μl
-Buffer 4 --> 3μl
-BSA -------> 1μl
-DNA ------> 10μl
-ECOR1 -----> 1.5μl
-PST1 ------> 1.5μl
- Restrictions were incubated at 37°C ON (overnight) and labeled as luc or mluc No. R-> ECO y PST Mar. and date.
- Melvin is also helping with the experiment, he did the same as me but for the Promega® click beetle luciferases, cutting them with XBAI and PSTI.
- I also inoculated 3 strains containing luciferase from photinus pyralis in 4ml of LB with 4μl of kanamycin and leave them growing at 37°C ON.