GSM1530468: NH-11 normal heart; Homo sapiens; MBD-Seq
Genomic DNA from cardiac samples was extracted using Phenol:Chloroform:Isoamyl Alcohol (25:24:1) and treated with RNase A (Invitrogen) according to manufacturer’s instructions. DNA fragmentation was performed on a Covaris S3 to obtain fragments with an average length of 200 bp. The fragment distribution was examined on a high sensitivity DNA chip. Methylated DNA was captured using the MethylCap kit (Diagenode AF-100-0048, Belgium). The concentrations of the fragmented and captured DNA were determined on a Fluostar Optima plate reader (BMG Labtech, Offenburg, Germany) with the Quant-iTTN Picogreen dsDNA assay kit (Invitrogen P7589, Merelbeke, Belgium). Library preparation and amplification were performed according to the ‘multiplexed paired-end ChIP protocol’ (with modifications) including the indexes from the ‘Multiplexing Sample Preparation Oligo Kit’ (Illumina, San Diego, California, USA). The libraries were 2 x 51 bp paired-end sequenced on an Illumina HiSeq 2000 platform. Base calling was performed with the Illumina Casava pipeline version 1.8.0. Initial sequencing quality assessment was based on data passing the Illumina Chastity filter.
GEO Accession
GSM1530468