User:Mariana Ruiz Velasco L./Notebook/IGEM 2010/Wet lab journal/2010/07/29

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When work takes over...
    1. lumP with XBAI and PST I stored as lumP 1 R-> XBA PST Mar and the date.  -H2O ---> 8μl -Buffer 3 --> 3μl -BSA --> 1μl -DNA ---> 15μl -XBA I -> 1.5μl -PST I --> 1.5μl  2. lumP with XBA I stored as lumP 1 R-> XBA Mar and the date.  -H2O ---> 11μl<br/ > -Buffer 4 --> 2μl<br/ > -BSA --> 1μl<br/ > -DNA ---> 5μl<br/ > -XBA I -> 1μl<br/ > <br/ > 3. lumP with PST I stored as lumP 1 R-> PST Mar and the date. <br/ > -H2O ---> 11μl<br/ > -Buffer 3 --> 2μl<br/ > -BSA --> 1μl<br/ > -DNA ---> 5μl<br/ > -PST I -> 1μl<br/ > <br/ > 4. Green luciferase with ECO RI and PST I stored as luc wt R-> ECO y PST Mar and the date.<br/ > <br/ > -H2O ---> 13μl<br/ > -Buffer 3 --> 3μl<br/ > -BSA --> 1μl<br/ > -DNA ---> 10μl<br/ > -ECO I -> 1.5μl<br/ > -PST I --> 1.5μl<br/ > <br/ > 5. Mutated-luciferase with ECO RI and PST I stored as mut-luz R-> ECO y PST Mar and the date. <br/ > -H2O ---> 13μl<br/ > -Buffer 3 --> 3μl<br/ > -BSA --> 1μl<br/ > -DNA ---> 10μl<br/ > -ECO RI -> 1.5μl<br/ > -PST I ---> 1.5μl<br/ > <br/ > 6. Plasmid PSB1C3 with each enzyme where ECO RI (buffer 2), XBA I (buffer 4), SPE I (buffer 4), and PST I (buffer 3) stored as p17 R-> enzyme Mar and the date. <br/ > -H2O ---> 11μl<br/ > -Buffer -> 2μl<br/ > -BSA --> 1μl<br/ > -DNA ---> 5μl<br/ > -Enzyme > 1μl<br/ > <br/ > 7. J23100 with SPE I and PST I stored as J23100 R-> SPE y PST Mar and the date. <br/ > -H2O ---> 8μl<br/ > -Buffer 2 --> 3μl<br/ > -BSA --> 1μl<br/ > -DNA ---> 15μl<br/ > -SPE I > 1.5μl<br/ > -PST I > 1.5μl<br/ > <br/ > 8. J23100 with each enzyme where ECO RI (buffer 2), XBA I (buffer 4), SPE I (buffer 4), and PST I (buffer 3). <br/ > -H2O ---> 11μl<br/ > -Buffer -> 2μl<br/ > -BSA --> 1μl<br/ > -DNA ---> 5μl<br/ > -Enzyme > 1μl<br/ > <br/ >
 * As soon as I arrived to the lab, I checked that my strains were growing well. I inoculated them in 4ml of LB with their respective antibiotic for ~7hr.<br/ >
 * Next, I extracted plasmid with the High Pure Plasmid Isolation Kit of Roche. Products were stored as p17/J23100/lumP(1-4) Mar Plasm Pur and the date respectively.<br/ >
 * Finally, I did the following restrictions so that tomorrow I am ready to ligate. <br/ >
 * Restrictions were incubated at 37°C ON.


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