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. 2014 Nov 25;15(1):1022. doi: 10.1186/1471-2164-15-1022

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© Pathak et al.; licensee BioMed Central Ltd. 2014

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Isolation and validation of Drosophila embryo MARs. (A) Overview of MAR DNA isolation procedure. (B) Size distribution of MAR DNA after electrophoresis on 1.2% agarose gel. M-Molecular weight marker; Lane 1-Genomic DNA; Lane 2-MAR DNA; Lane 3-MAR DNA + DNaseI; Lane 4-MAR DNA + RNaseA. (C) Slot-blot hybridization to show enrichment of known MARs in the MAR DNA preparation. Equal quantity of plasmids (1 μg) carrying sequences of BEAF protein exon as –ve control (slot i), MAR at histone gene locus (slot ii) and scs’ MAR at Hsp70 locus (slot iii) were loaded in each slot. Genomic DNA and MAR DNA were ³²P labelled by random primer labelling method and used for hybridization of the upper and the lower panel respectively. (D) Southern validation for 21 MARs and 8 non-MAR regions chosen on the basis of sequencing data. The upper panel shows the EtBr stained gel profile of the PCR amplified fragments and lower panel shows the blot probed with ³²P labelled MAR DNA.