Portal de Periódicos da CAPES

Bisulfite Sequencing of DNA

2010; Wiley; Volume: 91; Issue: 1 Linguagem: Inglês

10.1002/0471142727.mb0709s91

1934-3639

Russell P. Darst, Carolina Pardo‐Díaz, Lingbao Ai, Kevin D. Brown, Michael P. Kladde,

Cancer-related gene regulation

Abstract Exact positions of 5‐methylcytosine (m 5 C) on a single strand of DNA can be determined by bisulfite genomic sequencing (BGS). Treatment with bisulfite ion preferentially deaminates unmethylated cytosines, which are then converted to uracil upon desulfonation. Amplifying regions of interest from deaminated DNA and sequencing products cloned from amplicons permits determination of methylation at single‐nucleotide resolution along single DNA molecules, which is not possible with other methylation analysis techniques. This unit describes a BGS technique suitable for most DNA sources, including formaldehyde‐fixed tissue. Considerations for experimental design and common sources of error are discussed. Curr. Protoc. Mol. Biol . 91:7.9.1‐7.9.17. © 2010 by John Wiley & Sons, Inc.