User:Mariana Ruiz Velasco L./Notebook/IGEM 2010/Wet lab journal/2010/04/15

WiFi coli: A communicolight system

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First restriction :D I realized a double restriction using ECO RI-HF and PST I for a total of 20μl as follows: -H2O -------> 10μl -Buffer 2 --> 2μl -DNA -------> 5μl -ECO RI ---> 1μl -PSTI ------> 1μl Restrictions were incubated at 37°C ON(overnight). Defosfating restrictions Continuing with the experiment, I defosfated restrictions of plasmids 17, 18 (jorge), 18 and 30. The reaction was as follows for a total of 30μL. -H20 -------------> 11μL -Buffer ----------> 3μL -DNA ------------> 15μL -Defosfatase --> 1μL The reaction was incubated at 37°C for 15 min. and at 65°C for 5 min to inactivate the enzyme.