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Methods Mol Biol. 2018;1704:243-260. doi: 10.1007/978-1-4939-7463-4_8.

Comparative Metagenomics.

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Department of Biochemistry, Institute of Chemistry , University of São Paulo, São Paulo, SP, Brazil.
Medical Genomics Laboratory, CIPE/A.C. Camargo Cancer Center, São Paulo, SP, Brazil.
Instituto Federal de Alagoas, Maceió, Alagoas, Brazil.
Lab. of Neurosciences (LIM-27) Alzira Denise Hertzog Silva, Institute of Psychiatry, Faculdade de Medicina, Universidade de São Paulo (USP), São Paulo, SP, Brazil.
Department of Biochemistry, Institute of Chemistry, University of São Paulo, Av. Prof. Lineu Prestes, 748 room 909, 05508-000, São Paulo, SP, Brazil.


Thanks in large part to newer, better, and cheaper DNA sequencing technologies, an enormous number of metagenomic sequence datasets have been and continue to be generated, covering a huge variety of environmental niches, including several different human body sites. Comparing these metagenomes and identifying their commonalities and differences is a challenging task, due not only to the large amounts of data, but also because there are several methodological considerations that need to be taken into account to ensure an appropriate and sound comparison between datasets. In this chapter, we describe current techniques aimed at comparing metagenomes generated by 16S ribosomal RNA and shotgun DNA sequencing, emphasizing methodological issues that arise in these comparative studies. We provide a detailed case study to illustrate some of these techniques using data from the Human Microbiome Project comparing the microbial communities from ten buccal mucosa samples with ten tongue dorsum samples in terms of alpha diversity, beta diversity, and their taxonomic and functional profiles.


16S rRNA; Comparative metagenomics; DNA shotgun; Metagenome; Microbiome

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