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Methods Mol Biol. 2018;1746:63-75. doi: 10.1007/978-1-4939-7683-6_6.

Monolith Chromatography as Sample Preparation Step in Virome Studies of Water Samples.

Author information

1
Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia. ion.gutierrez@nib.si.
2
Department of Biotechnology and Systems Biology, National Institute of Biology, Ljubljana, Slovenia.
3
Lek Pharmaceuticals d.d., Menges, Slovenia.
4
University of Nottingham, Nottingham, UK.

Abstract

Viruses exist in aquatic media and many of them use this media as transmission route. Next-generation sequencing (NGS) technologies have opened new doors in virus research, allowing also to reveal a hidden diversity of viral species in aquatic environments. Not surprisingly, many of the newly discovered viruses are found in environmental fresh and marine waters. One of the problems in virome research can be the low amount of viral nucleic acids present in the sample in contrast to the background ones (host, eukaryotic, prokaryotic, environmental). Therefore, virus enrichment prior to NGS is necessary in many cases. In water samples, an added problem resides in the low concentration of viruses typically present in aquatic media. Different concentration strategies have been used to overcome such limitations. CIM monoliths are a new generation of chromatographic supports that due to their particular structural characteristics are very efficient in concentration and purification of viruses. In this chapter, we describe the use of CIM monolithic chromatography for sample preparation step in NGS studies targeting viruses in fresh or marine water. The step-by-step protocol will include a case study where CIM concentration was used to study the virome of a wastewater sample using NGS.

KEYWORDS:

CIM; Concentration; Monolithic chromatography; NGS; Water virome

PMID:
29492887
DOI:
10.1007/978-1-4939-7683-6_6
[Indexed for MEDLINE]

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