title: Analysis of Viral Epigenotypes Using Bisulfite Sequencing: A Detailed Protocol for the Crucial Bisulfite Modification and PCR Amplification Steps.
abstract: Characterization of viral DNA methylation patterns is essential to understand its function in viral pathogenesis. Bisulfite modification, followed by polymerase chain reaction (PCR) and sequencing is the most effective method for the high resolution methylation mapping of viral genomes. Since the bisulfite modification and PCR steps are the most critical ones, an optimized protocol for these two steps is presented, with special attention to potential pitfalls.
authors.affiliation: Department of Microbiology, Tumor and Cell Biology, Karolinska Institutet, Nobels väg 16, SE-171 77, Stockholm, Sweden. Daniel.salamon@ki.se.
authors.forename: Daniel
authors.initials: D
authors.lastname: Salamon
mesh_terms: D019175:DNA Methylation; D044127:Epigenesis, Genetic; D057890:Epigenomics / Q000379:methods; D016679:Genome, Viral; D005838:Genotype; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA; D013447:Sulfites
chemical_list: D013447:Sulfites; C042345:hydrogen sulfite
keywords: Bisulfite modification; DNA methylation; Polymerase chain reaction; Primer design; Sequencing; Virus