Request a detailed protocol DNA hydrolysis activity of varying time was assayed by the following: The reaction mixture contained 100 nM WT I-PpoI, 50 mM NaCl, 100 mM Tris (pH7.5), 1.5 mM dithiothreitol (DTT), 0.05 mg/ml bovine serum albumin (BSA), 50 nM DNA, 10 µM ethylenediaminetetraacetic acid (EDTA), and 4% glycerol. The hydrolysis assays were executed using a palindromic 5′-fluorescein-labeled DNA duplex (5′-TTG ACT CTC TTA AGA GAG TCA-3′).