User:Mariana Ruiz Velasco L./Notebook/IGEM 2010/Wet lab journal/2010/06/16

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 * style="background-color: #EEE"|[[Image:Logo_light_minimal_full_res.jpg|150px]] WiFi coli: A communicolight system
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Getting ready for punctual mutation
   -H2O ---> 18μl
 * Today, I inoculated E. coli strains containing a plasmid with a | constitutive promoter so that, after 6-7 hrs of incubation at 37°C, I can extract the plasmid and ligate the luciferase and double terminator with it.
 * I also did a restriction with Xba I and Pst I for a total of 30μL as follows:

-Buffer 2 --> 4μl

-BSA --> 1μl

-DNA ---> 5μl

-XBA I -> 1μl

-PST I --> 1μl   
 * Restrictions were incubated at 37°C for 4 hr and labeled as: lucif + TT c/rbs + 1 purif restr. XBA y PST Mar and the date in the top of the eppendorf. 
 * I also ran a gel at 90V for 50min to quantify the ligation PCR product with the mutated RBS to know how to prepare the mix for the PCR with Pfx.