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2 in dry methanol at room temperature for 36 h. The resulting metallated polymer was washed thoroughly with methanol (5×20 mL). From the AAS analysis, it was found that the amount of zinc present per gram polymer is equal to 11.5 mg.
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1542293182
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note
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DNA cleavage: Cleavage reaction was performed by suspending 100 μg of CPPL-Zn (corresponding to 0.88 mM of zinc if the polymer was to be completely soluble in buffer) in sodium cacodylate buffer (8 mM, 20 μL, pH 7.5, 35°C), containing pBR322 supercoiled plasmid DNA (10 ng/μL, New England Biolabs). 4 μL of methanol was used for polymer wetting. Individual reactions were quenched by adding gel loading buffer (5 μL) containing EDTA (100 mM) at regural time intervals. Reactions were loaded onto 0.7% agarose gel, containing ethidium bromide (1 μg/mL), and were electrophoresed for 1 h at constant current (80 mA). Gels were imaged on Bio-Rad Gel Documentation System 2000.
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49
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1542263360
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Lysozyme cleavage: CPPL-Zn (200 μg) was suspended in 20 μL of 10 mM cacodylate buffer (pH 7.5) containing lysozyme (25 μM) at 35°C. All reactions were quenched with 20 μL of sample buffer containing 20 μg of bromophenol blue and 20% of glycerol in 0.01 M of Tris-HCl buffer. Samples were loaded onto polyacrylamide gel (concentrating gel 4.5% and separating gel 12.5%) and electrophoresed for 8 h at constant voltage (80 V) in Tris-glycine buffer containing 0.1% sodium dodecyl sulphate. Gel was stained with Coomassie blue and destained prior to imaging on Bio-Rad Gel Doc 2000 system.
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