Extraction and purification of nucleic acids from viruses
Full Citation: Steward, G. F., and A. I. Culley. 2010. Extraction and purification of nucleic acids from viruses, p. 154-165. In S. W. Wilhelm, M. G. Weinbauer, and C. A. Suttle [eds.], Manual of Aquatic Viral Ecology. ASLO. [DOI 10.4319/mave.2010.978-0-9845591-0-7.154]
ABSTRACT: Research on the diversity and ecology of viruses in the environment has been revolutionized by the ability to detect, fingerprint, and sequence viral genes and genomes. The starting point for these molecular assays is the release and recovery of the viral nucleic acids. The complexity of this task depends in large part on the nature of the starting material and the purity and quality of the nucleic acids one requires for downstream applications. In some cases, simply heating the sample will suffice; in other cases, a series of organic extractions and purification in a buoyant density gradient may be required to achieve adequate purity. Our goal in this chapter is to assist the reader in making informed choices from among the many options available. Toward this end, we briefly review the methods that have been used to harvest and store viruses in preparation for extraction, and the methods by which their nucleic acids may be released and purified. We discuss the general principles upon which various commercial extraction kits are based and conclude with the presentation of four step-by-step protocols. We discuss the advantages and disadvantages of these protocols, and the ways in which they may be adapted to various situations.