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==Results== After running both enzymes through size-selection gels, it is clear that the Nb.BsrDI enzyme is '''NOT''' working properly: [[image:Nt.A1wI_Nb.BsrDI_test.jpg]] *Concluded, after checking the theoretical matches, there was an ''experimental'' error. It was determined that the Nb.BsrDI enzyme needed NEBuffer 2 before being incubated, which I failed to do. After testing the Nb.BsrDI enzyme with buffer, the results were clear: [[image:Nb.BsrDI_test.jpg]] Note: Gel appears to have a problem, but is sufficient for examining effects of enzyme Nb.BsrDI Clearly both enzymes work, the problem was in the experimental setup. After review of the NEB website on the activity of the enzymes I re-calibrated the numbers: For both enzymes, 1 unit can digest 1ug of DNA -> First measure amount of DNA via NanoDrop -> If necessary, split sample into 2 tubes and fill up to 50ul with water -> add 6ul 10x NEBuffer 2 -> add 2ul Nt.A1wI (or Nb.BsrDI) at 10U/ul -> incubate at correct temperature (37C or 65C, respectively) for 2 hours -> hold at 4C
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