MeCP2__Global_distribution_and_transcription. MeCP2__Global_distribution_and_transcription

MeCP2 is a nuclear protein with an affinity for methylated DNA that can recruit histone deacetylases. Deficiency or excess of MeCP2 causes severe neurological problems, highlighting the functional importance of this protein. Chromatin immunoprecipitation of MeCP2 in combination with high-throughput solexa sequencing identified genome-wide binding which tracks methyl-CpG density but does not grossly affect the binding of RNA polymerase II. Protocol: Brains were manually homogenised to a single cells suspension and crosslinked using 1% formaldehyde. Chromatin was sheared using a combination of Micrococcal nuclease and sonication (Skene et al, Mol Cell, 2010). Protein-DNA complexes were immunoprecipitated with the appropriate antibody and DNA was subsequently extracted. Adaptors were ligated to purified DNA which was then amplified by PCR. The purified DNA was captured on an Illumina flow cell for cluster generation. Libraries were sequenced on the Genome Analyzer following the manufacturer's protocols Sequencing was performed at the Wellcome Trust Sanger Institute; Hinxton, Cambridge, UK. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/