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From 2010.igem.org

1. Mix together the following:

    a. 5 ul deionized water
    b. 2 ul of Reaction Buffer
    c. 5 ul of digested downstream insert (or water if using only 1 insert)
    d. 5 ul of digested downstream insert 
    e. 3 ul of digested vector
    f. 1 ul of DNA ligase

2. Incubate at room-temp for 5 mins

3. Kill reaction for 20 mins at 80 degrees Celsius