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Next-generation Sequencing of the DNA Virome from Fecal Samples
Cynthia L Monaco1, Douglas S Kwon2,3
1Department of Medicine, Washington University School of Medicine, St. Louis, USA.
Abstract:
Herein we describe a detailed protocol for DNA virome analysis of low input human stool samples (Monaco et al., 2016). This protocol is divided into four main steps: 1) stool samples are pulverized to evenly distribute microbial matter; 2) stool is enriched for virus-like particles and DNA is extracted by phenol-chloroform; 3) purified DNA is multiple-strand displacement amplified (MDA) and fragmented; and 4) libraries are constructed and sequenced using Illumina Miseq. Subsequent sequence analysis for viral sequence identification should be sensitive but stringent.
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