title: Microbial Community Profiling Using Terminal Restriction Fragment Length Polymorphism (T-RFLP) and Denaturing Gradient Gel Electrophoresis (DGGE).
abstract: In their natural environments, microorganisms usually live in organized communities. Profiling analysis of microbial communities has recently assumed special relevance as it allows a thorough understanding of the diversity of the microbiota, its behavior over time, and the establishment of patterns associated with health and disease. The application of molecular biology approaches holds the advantage of including culture-difficult and as-yet-uncultivated phylotypes in the profiles, providing a more comprehensive picture of the microbial community. This chapter focuses on two particular techniques, namely, terminal restriction fragment length polymorphism (T-RFLP) and denaturing gradient gel electrophoresis (DGGE), both of which have been widely used in environmental studies and have been successfully used by the authors in the study of the oral microbial communities associated with conditions of health and disease.
authors.affiliation: Molecular Microbiology Laboratory, Department of Endodontics, Estácio de Sá University, Rio de Janeiro, Brazil. jf_siqueira@yahoo.com.|Faculty of Dentistry, Estácio de Sá University, Av. Alfredo Baltazar da Silveira, 580/cobertura, Recreio, Rio de Janeiro, RJ, 22790-710, Brazil. jf_siqueira@yahoo.com.
authors.forename: José F
authors.initials: JF
authors.lastname: Siqueira
authors.affiliation: Microbe Division/Japan Collection of Microorganisms, RIKEN BioResource Center, Wako, Saitama, Japan.
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Sakamoto
authors.affiliation: Institute of Microbiology Prof. Paulo de Góes, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Alexandre S
authors.initials: AS
authors.lastname: Rosado
mesh_terms: D058645:Denaturing Gradient Gel Electrophoresis / Q000379:methods; D006801:Humans; D054892:Metagenome; D056186:Metagenomics / Q000379:methods; D064307:Microbiota; D009055:Mouth / Q000382:microbiology; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012336:RNA, Ribosomal, 16S / Q000235:genetics
chemical_list: D012336:RNA, Ribosomal, 16S
keywords: 16S rRNA gene; Denaturing gradient gel electrophoresis (DGGE); Human oral microbiota; Terminal restriction fragment length polymorphism (T-RFLP)