| Laboratory procedures to generate viral metagenomes. | |
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MedLine Citation:
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PMID: 19300441 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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This collection of laboratory protocols describes the steps to collect viruses from various samples with the specific aim of generating viral metagenome sequence libraries (viromes). Viral metagenomics, the study of uncultured viral nucleic acid sequences from different biomes, relies on several concentration, purification, extraction, sequencing and heuristic bioinformatic methods. No single technique can provide an all-inclusive approach, and therefore the protocols presented here will be discussed in terms of hypothetical projects. However, care must be taken to individualize each step depending on the source and type of viral-particles. This protocol is a description of the processes we have successfully used to: (i) concentrate viral particles from various types of samples, (ii) eliminate contaminating cells and free nucleic acids and (iii) extract, amplify and purify viral nucleic acids. Overall, a sample can be processed to isolate viral nucleic acids suitable for high-throughput sequencing in approximately 1 week. |
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Authors:
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Rebecca V Thurber; Matthew Haynes; Mya Breitbart; Linda Wegley; Forest Rohwer |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S. |
Journal Detail:
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Title: Nature protocols Volume: 4 ISSN: 1750-2799 ISO Abbreviation: Nat Protoc Publication Date: 2009 |
Date Detail:
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Created Date: 2009-03-20 Completed Date: 2009-06-12 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101284307 Medline TA: Nat Protoc Country: England |
Other Details:
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Languages: eng Pagination: 470-83 Citation Subset: IM |
Affiliation:
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Department of Biology, San Diego State University, San Diego, California 92182, USA. rvegathurber@gmail.com |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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DNA Viruses
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genetics*,
isolation & purification Genome, Viral* Genomic Library* Genomics / methods* Nucleic Acid Amplification Techniques Ultracentrifugation Virion / genetics, isolation & purification Virus Cultivation |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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