User:Mariana Ruiz Velasco L./Notebook/IGEM 2010/Wet lab journal/2010/06/07

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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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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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Ligation day!!!
    -H2O ---> 6μL -Buffer ligase ---> 2μL -Vector (plasmid) -> 3μL -Insert (luciferase) ---> 3μL -Insert (double ter.) --> 5μL -Ligase > 1μL   -H2O ---> 7μL -Buffer ligase ---> 2μL -Vector (plasmid) -> 2μL<br/ > -Insert (luciferase) ---> 3μL<br/ > -Insert (double ter.) --> 5μL<br/ > -Ligase > 1μL<br/ > <br/ > <br/ > -H2O ---> 7μL<br/ > -Buffer ligase ---> 2μL<br/ > -Vector (plasmid) -> 2μL<br/ > -Insert (luciferase) ---> 3μL<br/ > -Insert (double ter.) --> 5μL<br/ > -Ligase > 1μL<br/ > <br/ > <br/ > -H2O ---> 7μL<br/ > -Buffer ligase ---> 2μL<br/ > -Vector (plasmid) -> 2μL<br/ > -Insert (luciferase) ---> 3μL<br/ > -Insert (double ter.) --> 5μL<br/ > -Ligase > 1μL<br/ > <br/ > <br/ >
 * As in the previous days there was no success in the ligations, I did four with different combinations in order to optimize the probabilities of getting colonies transformed with both, luciferase and the double terminator.
 * The ligations were done for a total of 20μL, using different restriction enzymes and either the luciferase plasmid or the purified luciferase PCR product, and a double terminator (TT) amplified by PCR of the plasmid with the TT. All ligations were labeled as lig. 1-4 TAA M, ligación lucif + TT Mar
 * 1: luciferase plasmid + TT PCR (1-F)
 * 2: luciferase plasmid + TT plasmid
 * 3: luciferase PCR + TT PCR
 * 4: luciferase PCR + TT plasmid
 * After the ligation, I transformed competent cells and plated them in tetracycline-LB plates.


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