Bisulfite Sequencing of Homo Sapiens
Library synthesis: Peripheral blood mononuclear cells were obtained from patient blood samples by ficoll-hypaque (GE Healthcare) gradient. Dynabeads CD4 Positive Isolation Kit (Thermo Fisher Scientific, MA, USA) was used to separate CD4+ cells. Genomic DNA was isolated from CD4+ cells using DNeasy Mini Kit (Qiagen, Carlsbad, CA). All protocols were made according to manufacturers procedures. Libraries were generated using the SureSelectXT Human Methyl-Seq (Agilent) Targeted Enrichment Kit (Agilent Technologies, Santa Clara, CA). Combining the SureSelectXT system and bisulfite conversion, studying 80Mb and covering over 3.7 million CpGs. Samples were processed following the recommended MethylSeq protocol Version B, January 2013. The samples target sequences were bisulfite converted using the EZ DNA MethylationGold kit (Zymo Research) as described in the MethylSeq protocol.