ERX032776 SAMEA1094599 Transcription_profiling_by_high_throughput_sequencing_of_long_non-coding_RNAs_across_multiple_rodent_species ChIP-Seq GENOMIC ChIP 0 Application_Read Forward 1 Illumina_Genome_Analyzer_II ERA070557 ERS074665 ERP001022 PRJEB2764 2014-08-12T15:38:14Z Organism taxonomy_id="10116" taxonomy_name="Rattus norvegicus" Alias=E-MTAB-867:rno8 Broker name=ArrayExpress Description=Protocols: Samples were prepared by direct perfusion of the tissue with buffered salt solution, followed by % formaldehyde. After 10 minutes, the tissue was removed and diced in 500 mM glycine buffer to neutralize the formaldehyde. After homogenization, the cells were rinsed with PBS. The animal tissues were crosslinked with formaldehyde treatment and chromatin fragmented to an average of 300 bp by sonication. Chromatin from an mass equivalent of 1/4 mouse liver was used for each ChIP experiment. Solexa libraries were prepared following the instructions of Illumina (Sample preparation for genomic DNA - version 2.2) with the following modifications. The ChIP-enriched DNA was not further fragmented. After end-repair and addition of an A base to the 3prime ends, the adapters were ligated to the ends of the DNA Fragments using 2 ul of fortyfold diluted 'Adapter oligo mix' in a total reaction volume of 25 ul. Between these steps, the DNA was purified using the DNA Clean-and-Concentrator-5 kit (Zymo Research). Subsequently, the DNA was amplified by 18 cycles of PCR, purified with QIAquick PCR purification Kit, and eluted with 33.5 ul of 10 mM tris buffer at pH7.0. The PCR-product was sized fractionated on 2% agarose gel and a gel slice containing the 200-300 bp fragments was excised. The flowcells were prepared and processed according to the manufacturer's protocols, with single-end sequencing for 36 to 45 cycles. DevelopmentalStage=adult INSDC center alias=CRI INSDC center name=Cancer Research UK Cambridge Research Institute INSDC first public=2012-04-01T17:00:37Z INSDC last update=2018-03-08T15:33:19Z INSDC status=public OrganismPart=liver SRA accession=ERS074665 Sample Name=ERS074665 Sex=male Title=rno8