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AFLP protocol comparison for microbial diversity fingerprinting.
Bertani, Gaia; Savo Sardaro, Maria Luisa; Neviani, Erasmo; Lazzi, Camilla.
Afiliação
  • Bertani G; Food and Drug Department, University of Parma, Parco Area delle Scienze 49/A, Parma, Italy.
  • Savo Sardaro ML; Food and Drug Department, University of Parma, Parco Area delle Scienze 49/A, Parma, Italy. marialuisa.savosardaro@unisanranffaele.gov.it.
  • Neviani E; Department of Human Science and Promotion of the Quality of Life, University San Raffaele Srl, Via Val Cannuta 247, Rome, Italy. marialuisa.savosardaro@unisanranffaele.gov.it.
  • Lazzi C; Food and Drug Department, University of Parma, Parco Area delle Scienze 49/A, Parma, Italy.
J Appl Genet ; 60(2): 217-223, 2019 May.
Article em En | MEDLINE | ID: mdl-30989627
Over the last decade, several methods based on genomic DNA have been developed for the identification and genotyping of prokaryotic and eukaryotic organisms. These genomic methods differ regarding taxonomic range, discriminatory power, reproducibility, and ease of interpretation and standardization. The amplified fragment length polymorphism (AFLP) technique is a very powerful DNA fingerprinting technique for DNA of any source or complexity, varying in both size and base composition. In addition, this method shows high discriminatory power and good reproducibility allowing it to be efficient in discriminating at both the species and strain levels. The development and application of AFLP have allowed significant progress in the study of biodiversity and taxonomy of microorganisms. In the last years, the Applied Biosystems AFLP Microbial Fingerprinting Kit, now out of production, was widely used in various studies to perform AFLP characterization of selected bacteria strains (described by Vos et al. (Nucleic Acids Res 23(21):4407-4414, 1995)). Its replacement gives the possibility for laboratories to continue the use of the previous AFLP data as a reference for bacteria genetic fingerprinting analysis in biodiversity studies. To overcome this issue a result comparison, by using an improved AFLP protocol and the AFLP commercial kit, was performed. In particular, previous results on different species (Listeria monocytogenes, Lactobacillus plantarum, and Streptococcus thermophilus) obtained with the commercial kit were compared with the improved AFLP procedure to validate the protocol. When compared with the AFLP Microbial Fingerprinting Kit, the improved protocol shows high reproducibility, resolution, and overall, is a faster method with lower costs.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Bactérias / DNA Bacteriano / Impressões Digitais de DNA / Análise do Polimorfismo de Comprimento de Fragmentos Amplificados Idioma: En Ano de publicação: 2019 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Bactérias / DNA Bacteriano / Impressões Digitais de DNA / Análise do Polimorfismo de Comprimento de Fragmentos Amplificados Idioma: En Ano de publicação: 2019 Tipo de documento: Article