Optimization and confirmation of human chromatin assembly reaction. a DNA supercoiling assay to optimize the ratio of HsH2A/HsH2B and HsH3.1/HsH4 mRNAs in the chromatin reconstitution reaction for 4 h. b Supercoiling assay of assembled human chromatin. Incubation time indicated with 0, 1, 4, 6, and 8 h, where 0 indicated the reconstitution reaction in the absence of mRNAs encoding histones. Supercoils were analyzed by 0.8% agarose <t>TBE</t> gel. Relaxed, linear, and <t>supercoiled</t> DNA are indicated as RC, L, and SC, respectively. c Partial MNase digestion of assembled human chromatin, then samples were run on 2.0% agarose gel. Reconstituted chromatin was digested by MNase for the indicated time (0, 5, 10, 20, 30, and 40 min), and bands corresponding to digestion fragment of the mono-, di-, tri- and tetranucleosome were indicated. Mr: DNA molecular weight marker
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