Extraction and purification of viruses from fecal samples for metagenome and morphology analyses
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Extraction and purification of viruses from fecal samples for metagenome and morphology analyses. / Castro-Mejía, Josué L.; Deng, Ling; Vogensen, Finn K.; Reyes, Alejandro; Nielsen, Dennis S.
The Human Virome: Methods and Protocols. ed. / Andrés Moya; Vicente Pérez Brocal. Humana Press, 2018. p. 49-57 (Methods in Molecular Biology, Vol. 1838).Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
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TY - CHAP
T1 - Extraction and purification of viruses from fecal samples for metagenome and morphology analyses
AU - Castro-Mejía, Josué L.
AU - Deng, Ling
AU - Vogensen, Finn K.
AU - Reyes, Alejandro
AU - Nielsen, Dennis S.
PY - 2018
Y1 - 2018
N2 - The human enteric virome consists of endogenous retro elements and viruses that infect the host and members of the gut microbiome (GM). Mounting evidence suggests that the gut virome plays a central role in maintaining homeostasis and via the GM influences immunology of the host. To thoroughly characterize the gut virome, it is often very useful to first separate and concentrate extracellular viral-like particles (eVLPs) enabling an integrative characterization of them. Here, we describe a detailed protocol for extraction and concentration of the viral fraction from fecal samples based on a polyethylene glycol precipitation (PEG) approach. These procedures maximize the yields of eVLPs (and their DNA) with high purity well suited for down-stream analysis such as quantification and morphological assessment, determination of phage-host pairs as well as virome sequencing.
AB - The human enteric virome consists of endogenous retro elements and viruses that infect the host and members of the gut microbiome (GM). Mounting evidence suggests that the gut virome plays a central role in maintaining homeostasis and via the GM influences immunology of the host. To thoroughly characterize the gut virome, it is often very useful to first separate and concentrate extracellular viral-like particles (eVLPs) enabling an integrative characterization of them. Here, we describe a detailed protocol for extraction and concentration of the viral fraction from fecal samples based on a polyethylene glycol precipitation (PEG) approach. These procedures maximize the yields of eVLPs (and their DNA) with high purity well suited for down-stream analysis such as quantification and morphological assessment, determination of phage-host pairs as well as virome sequencing.
KW - Extraction procedures
KW - Gut-virome
KW - Metagenome of extracellular viral-like particles (eVLPs)
KW - Morphology
KW - Polyethylene-glycol (PEG) precipitation
U2 - 10.1007/978-1-4939-8682-8_5
DO - 10.1007/978-1-4939-8682-8_5
M3 - Book chapter
C2 - 30128989
AN - SCOPUS:85052194015
SN - 978-1-4939-8681-1
T3 - Methods in Molecular Biology
SP - 49
EP - 57
BT - The Human Virome
A2 - Moya, Andrés
A2 - Brocal, Vicente Pérez
PB - Humana Press
ER -
ID: 210012489