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Fig. 3 | BMC Biotechnology

Fig. 3

From: Reconstitution of Drosophila and human chromatins by wheat germ cell-free co-expression system

Fig. 3

DNA supercoiling and MNase assays of Drosophila chromatin assembly reactions. a Determination of the appropriate ratio of DmH2A/DmH2B and DmH3/DmH4 mRNAs in the Drosophila chromatin reconstitution reaction. The incubation time was 4 h. Supercoiling assay was performed, samples were analyzed by 0.8% agarose TBE gel. Relaxed, linear, and supercoiled plasmid DNAs are indicated as RC, L, and SC, respectively. b Supercoiling assay of assembled Drosophila chromatin under the indicated reaction time 0, 1, 4, and 6 h, where 0 indicated the reconstitution reaction in the absence of mRNAs encoding histones. Supercoiling assay was performed, samples were analyzed by 0.8% agarose TBE gel. c Partial MNase digestion was performed on assembled Drosophila chromatin for the indicated time (0, 5, 10, 20, 30, and 40 min), then samples were run on 2.0% agarose gel. Bands corresponding to digestion fragment of the mono-, di-, and trinucleosomes were detected and indicated. Agarose gels were visualized by ethidium bromide and UV light. Mr: DNA molecular weight marker

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