Normalization of environmental metagenomic DNA enhances the discovery of under-represented microbial community members
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Date
Authors
Ramond, Jean-Baptiste
Makhalanyane, Thulani Peter
Tuffin, Marla I.
Cowan, Don A.
Journal Title
Journal ISSN
Volume Title
Publisher
Wiley
Abstract
Normalization is a procedure classically employed to detect rare sequences in cellular
expression profiles (i.e. cDNA libraries). Here, we present a normalization protocol involving
the direct treatment of extracted environmental metagenomic DNA with S1 nuclease; referred
to as Normalization of metagenomic DNA: NmDNA. We demonstrate that NmDNA, prior to
post hoc PCR based experiments (16S rRNA gene T-RFLP fingerprinting and clone library),
increased the diversity of sequences retrieved from environmental microbial communities by
detection of rarer sequences. This approach could be used to enhance the resolution of
detection of ecologically relevant rare members in environmental microbial assemblages.
Description
Keywords
Normalization, Environmental metagenomic, DNA enhances, Discovery, Novel microbial diversity
Sustainable Development Goals
Citation
Ramond, J.-B, Makhalanyane, TP, Tuffin, MI & Cowan, DA 2015, 'Normalization of environmental metagenomic DNA enhances the discovery of under-represented microbial community members', Letters in Applied Microbiology, vol. 60, no. 4, pp. 359-366.