Instrument: Illumina Genome Analyzer II
Strategy: Bisulfite-Seq
Source: GENOMIC
Selection: Reduced Representation
Layout: SINGLE
Construction protocol: Genomic DNA from sorted cell fractions was isolated with the DNeasy Blood & Tissue Kit (Qiagen, Hilden, Germany). Genomic DNA was digested with the methylation sensitive restriction enzyme MspI (New England BioLabs, Frankfurt a. M., Germany). End repair of the resulting 3' overhangs and A-tailing were performed to enable adaptor ligation. After purification with AMPure XP beads (Beckmann Coulter, Krefeld, Germany) and ligation of the methylated universal adaptors (Illumina, San Diego, USA), an additional bead wash step was conducted. Bisulfite conversion was performed with the EpiTect Bisulfite Kit (Qiagen, Hilden, Germany), followed by a wash step using the MinElute PCR Purification Kit (Qiagen). After barcoding of the bisulfite treated samples two additional rounds of bead clean-up were performed. The final libraries were sequenced (single end 76 bp) with the Illumina Genome Analyzer II (Illumina, San Diego, USA) according to the manufacturer's instructions.