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Assessing the Fecal Microbiota: An Optimized Ion Torrent 16S rRNA Gene-Based Analysis Protocol

2013; Public Library of Science; Volume: 8; Issue: 7 Linguagem: Inglês

10.1371/journal.pone.0068739

1932-6203

Christian Milani, Arancha Hevia, Elena Foroni, Sabrina Duranti, Francesca Turroni, Gabriele Andrea Lugli, Borja Sánchez, Rebeca Martín, Miguel Gueimonde, Douwe van Sinderen, Abelardo Margollés, Marco Ventura,

Fecal contamination and water quality

Assessing the distribution of 16S rRNA gene sequences within a biological sample represents the current state-of-the-art for determination of human gut microbiota composition. Advances in dissecting the microbial biodiversity of this ecosystem have very much been dependent on the development of novel high-throughput DNA sequencing technologies, like the Ion Torrent. However, the precise representation of this bacterial community may be affected by the protocols used for DNA extraction as well as by the PCR primers employed in the amplification reaction. Here, we describe an optimized protocol for 16S rRNA gene-based profiling of the fecal microbiota.