Analysis of a viral metagenomic library from 200 m depth in Monterey Bay, California constructed by direct shotgun cloning
We constructed a viral metagenome from a sample collected below the euphotic zone in a eutrophic bay of coastal California. We concentrated microbes from approximately one cubic meter of seawater collected from 200 m depth in Monterey Bay, CA using tangential flow filtration. Cellular material was depleted by differential centrifugation and filtration (0.2 ?m) and the viruses enriched, concetrated and purified by banding in a CsCl equilibrium buoyant density gradient. DNA was extracted from the virus fraction of the CsCl gradient, sheared, and cloned with no prior amplification into a plasmid vector and propagated in E. coli to produce the MBv200m library. Random clones were sequenced by the Sanger method. |  |  | |