Journal
METHODSX
Volume 1, Issue -, Pages 197-206Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.mex.2014.09.001
Keywords
Marine viruses; Concentration; Tangential flow filtration; Ultrafiltration; High turbidity seawater
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Funding
- National Natural Science Foundation of China [41376135, 81341082]
- Program for Professor of Special Appointment (Eastern Scholar) from Shanghai Municipal Education Commission [20101222]
- Doctoral Fund of Ministry of Education of China [20133104110006]
- Innovation Program of Shanghai Municipal Education Commission, China [14ZZ144]
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Marine viruses are the most abundant entities in the ocean and play crucial roles in the marine ecological system. However, understanding of viral diversity on large scale depends on efficient and reliable viral purification and concentration techniques. Here, we report on developing an efficient method to purify and concentrate viruses from large body of high turbidity seawater. The developed method characterizes with high viral recovery efficiency, high concentration factor, high viral particle densities and high-throughput, and is reliable for viral concentration from high turbidity seawater. Recovered viral particles were used directly for subsequent analysis by epifluorescence microscopy, transmission electron microscopy and metagenomic sequencing. Three points are essential for this method: center dot The sampled seawater (> 150L) was initially divided into two parts, water fraction and settled matter fraction, after natural sedimentation. center dot Both viruses in the water fraction concentrated by tangential flowfiltration (TFF) and viruses isolated from the settled matter fraction were considered as the whole viral community in high turbidity seawater. center dot The viral concentrates were re-concentrated by using centrifugal filter device in order to obtain high density of viral particles. (C) 2014 The Authors. Published by Elsevier B.V.
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