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

WiFi coli: A communi-colight system

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When work takes over... 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. 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. Finally, I did the following restrictions so that tomorrow I am ready to ligate. 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 -Buffer 4 --> 2μl -BSA ------> 1μl -DNA -------> 5μl -XBA I -----> 1μl 3. lumP with PST I stored as lumP 1 R-> PST Mar and the date. -H2O -------> 11μl -Buffer 3 --> 2μl -BSA ------> 1μl -DNA -------> 5μl -PST I -----> 1μl 4. Green luciferase with ECO RI and PST I stored as luc wt R-> ECO y PST Mar and the date. -H2O -------> 13μl -Buffer 3 --> 3μl -BSA ------> 1μl -DNA -------> 10μl -ECO I -----> 1.5μl -PST I ------> 1.5μl 5. Mutated-luciferase with ECO RI and PST I stored as mut-luz R-> ECO y PST Mar and the date. -H2O -------> 13μl -Buffer 3 --> 3μl -BSA ------> 1μl -DNA -------> 10μl -ECO RI -----> 1.5μl -PST I -------> 1.5μl 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. -H2O -------> 11μl -Buffer -----> 2μl -BSA ------> 1μl -DNA -------> 5μl -Enzyme ----> 1μl 7. J23100 with SPE I and PST I stored as J23100 R-> SPE y PST Mar and the date. -H2O -------> 8μl -Buffer 2 --> 3μl -BSA ------> 1μl -DNA -------> 15μl -SPE I ----> 1.5μl -PST I ----> 1.5μl 8. J23100 with each enzyme where ECO RI (buffer 2), XBA I (buffer 4), SPE I (buffer 4), and PST I (buffer 3). -H2O -------> 11μl -Buffer -----> 2μl -BSA ------> 1μl -DNA -------> 5μl -Enzyme ----> 1μl Restrictions were incubated at 37°C ON.