Marine methylotrophs revealed by stable-isotope probing, multiple displacement amplification and metagenomics

Neufeld, JD, Chen, Y, Dumont, MG and Murrell, JC (2008) Marine methylotrophs revealed by stable-isotope probing, multiple displacement amplification and metagenomics. Environmental Microbiology, 10 (6). pp. 1526-1535. ISSN 1462-2920

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Abstract

The concentrations of one-carbon substrates that fuel methylotrophic microbial communities in the ocean are limited and the specialized guilds of bacteria that use these molecules may exist at low relative abundance. As a result, these organisms are difficult to identify and are often missed with existing cultivation and gene retrieval methods. Here, we demonstrate a novel proof of concept: using environmentally-relevant substrate concentrations in stable-isotope probing (SIP) incubations to yield sufficient DNA for large-insert metagenomic analysis through multiple displacement amplification (MDA). A marine surface-water sample was labelled sufficiently by incubation with near in situ concentrations of methanol. Picogram quantities of labelled (13)C-DNA were purified from caesium chloride gradients, amplified with MDA to produce microgram amounts of high-molecular-weight DNA (

Item Type: Article
Faculty \ School: Faculty of Science > School of Environmental Sciences
Related URLs:
Depositing User: Rhiannon Harvey
Date Deposited: 27 Mar 2012 15:15
Last Modified: 21 Apr 2020 17:53
URI: https://ueaeprints.uea.ac.uk/id/eprint/38517
DOI: 10.1111/j.1462-2920.2008.01568.x

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