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

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First restriction :D


 -H2O ---> 10μl -Buffer 2 --> 2μl -DNA ---> 5μl -ECO RI ---> 1μl -PSTI --> 1μl  
 * I realized a double restriction using ECO RI-HF and PST I for a total of 20μl as follows:
 * Restrictions were incubated at 37°C ON(overnight).

Defosfating restrictions
  -H20 -> 11μL -Buffer --> 3μL -DNA > 15μL -Defosfatase --> 1μL<br/ > <br/ >
 * 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.


 * The reaction was incubated at 37°C for 15 min. and at 65°C for 5 min to inactivate the enzyme.


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