title: Methyl-binding DNA capture Sequencing for Patient Tissues.
abstract: Methylation is one of the essential epigenetic modifications to the DNA, which is responsible for the precise regulation of genes required for stable development and differentiation of different tissue types. Dysregulation of this process is often the hallmark of various diseases like cancer. Here, we outline one of the recent sequencing techniques, Methyl-Binding DNA Capture sequencing (MBDCap-seq), used to quantify methylation in various normal and disease tissues for large patient cohorts. We describe a detailed protocol of this affinity enrichment approach along with a bioinformatics pipeline to achieve optimal quantification. This technique has been used to sequence hundreds of patients across various cancer types as a part of the 1,000 methylome project (Cancer Methylome System).
authors.affiliation: Department of Molecular Medicine, University of Texas Health Science Center at San Antonio.
authors.forename: Rohit R
authors.initials: RR
authors.lastname: Jadhav
authors.affiliation: Department of Molecular Medicine, University of Texas Health Science Center at San Antonio.
authors.forename: Yao V
authors.initials: YV
authors.lastname: Wang
authors.affiliation: Department of Molecular Medicine, University of Texas Health Science Center at San Antonio.
authors.forename: Ya-Ting
authors.initials: YT
authors.lastname: Hsu
authors.affiliation: Department of Molecular Medicine, University of Texas Health Science Center at San Antonio.
authors.forename: Joseph
authors.initials: J
authors.lastname: Liu
authors.affiliation: Greehey Children's Cancer Research Institute, University of Texas Health Science Center at San Antonio.
authors.forename: Dawn
authors.initials: D
authors.lastname: Garcia
authors.affiliation: Greehey Children's Cancer Research Institute, University of Texas Health Science Center at San Antonio.
authors.forename: Zhao
authors.initials: Z
authors.lastname: Lai
authors.affiliation: Department of Molecular Medicine, University of Texas Health Science Center at San Antonio.
authors.forename: Tim H M
authors.initials: TH
authors.lastname: Huang
authors.affiliation: Department of Molecular Medicine, University of Texas Health Science Center at San Antonio; jinv@uthscsa.edu.
authors.forename: Victor X
authors.initials: VX
authors.lastname: Jin
mesh_terms: D019295:Computational Biology / Q000379:methods*; D004247:DNA; D019175:DNA Methylation; D044127:Epigenesis, Genetic; D006801:Humans; D017422:Sequence Analysis, DNA
chemical_list: D004247:DNA