User:Mariana Ruiz Velasco L./Notebook/IGEM 2010/Wet lab journal/2010/05/31

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 * style="background-color: #EEE"|[[Image:Logo_light_minimal_full_res.jpg|150px]] WiFi coli: A communicolight system
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


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Ready for ligation!!!
 
 * What I did today was a restriction of the PCR purified product (luciferase) with ECO RI and SPEI for a total of 40μl and stored as ECO RI y SPE I Prod. luc. PCR Mar and date (1-4). The reaction is described next:

-H2O -> 18μl -BSA -> 1μl -Buffer --> 4μl -ECO RI -> 1μl -SPEI > 1μl -DNA > 15μl     1. Ladder 2. Blank 3. Double terminator | BBa_B0015<br/ > 4. Restriction of luciferase<br/ > 5. Negative control (water)<br/ > 6. Positive control (plasmid with luciferase but without restriction)<br/ > <br/ > <br/ > <br/ > -H2O --> 12μL<br/ > -Buffer ligase --> 2μL<br/ > -Vector (plasmid) > 2μL<br/ > -Insert (luciferase) --> 3μL<br/ > -Ligase ---> 1μL<br/ > <br/ > <br/ >
 * In order to quantify the vector-insert relation for the ligation, I ran a 2% agarose gel for 50 min at 90V:
 * The lanes are as follow:
 * Finally, I did the ligation as follows for a total of 20μL:
 * The mix was incubated at 22°C for 10 min and at 70°C for 10 min to inactivate the enzyme.


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