title: GSM2611382: 104_CS_S2_BO_12_B1; Vitis vinifera; Bisulfite-Seq
library_construction_protocol: Genomic DNA was isolated from approximately 100 mg of berry pericarps by the Doyle&Doyle protocol (Focus, 1990) Reduced Representation Bisulfite Sequencing (RRBS) libraries were constructed acccording to Gu et al. 2011 with modifications. DNA was digested with TaqI enzyme (recognition site T^CGA, NEB, Ipswich, MA, USA) at 65°C for 2 h. After purification using the QIAquick PCR Purification kit (QIAGEN, Germany), fragment ends were repaired and ligated using adapters and the protocol provided by the Ovation Ultralow Methyl-Seq DR Multiplex Kit (NuGEN, CA, USA). Ligated products corresponding to 100–1500 bp DNA fragments were purified by 2% Low-Range agarose gel electrophoresis. A final end-repair step of the purified fragments was performed using the same NuGEN kit. Bisulfite conversion was conducted using a EpiTect Fast DNA Bisulfite Kit (QIAGEN, Germany). The final RRBS libraries were generated by PCR amplification and validated by an Agilent 2100 Bioanalyzer (Agilent Technologies). Reduced Representation Bisulfite Sequencing (RRBS) libraries sequenced using the Illumina HiSeq2500 platform (Illumina, San Diego, CA, USA) in paired-ended (PE), 125 bp runs