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Detection and enumeration of Lak megaphages in microbiome samples by endpoint and quantitative PCR

Crisci, MA; Corsini, PM; Bordin, N; Chen, LX; Banfield, JF; Santini, JM; (2022) Detection and enumeration of Lak megaphages in microbiome samples by endpoint and quantitative PCR. STAR Protocols , 3 (1) 10.1016/j.xpro.2021.101029. Green open access

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Abstract

Lak megaphages are prevalent across diverse gut microbiomes and may potentially impact animal and human health through lysis of Prevotella. Given their large genome size (up to 660 kbp), Lak megaphages are difficult to culture, and their identification relies on molecular techniques. Here, we present optimized protocols for identifying Lak phages in various microbiome samples, including procedures for DNA extraction, followed by detection and quantification of genes encoding Lak structural proteins using diagnostic endpoint and SYBR green-based quantitative PCR, respectively. For complete details on the use and execution of this protocol, please refer to Crisci et al., (2021).

Type: Article
Title: Detection and enumeration of Lak megaphages in microbiome samples by endpoint and quantitative PCR
Open access status: An open access version is available from UCL Discovery
DOI: 10.1016/j.xpro.2021.101029
Publisher version: https://doi.org/10.1016/j.xpro.2021.101029
Language: English
Additional information: © 2021 The Author(s). This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Structural and Molecular Biology
URI: https://discovery.ucl.ac.uk/id/eprint/10141284
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