Abstract:
This protocol outlines the use of multiple displacement amplification (MDA) as a form of whole-genome amplification (WGA), with application to a number of different analytical techniques in environmental microbiology. The MDA reaction uses the phi29 enzyme to amplify low concentrations of nucleic acid for subsequent characterization. This approach enables access to low-abundance DNA from low-biomass samples that would otherwise be recalcitrant to analysis with tools such as polymerase chain reaction (PCR), microarray analysis, metagenomics, metatranscriptomics, and single-cell cultivation. This efficient form of WGA has provided microbial ecologists with a means to investigate novel and challenging environments, as well as providing solutions for novel single-cell applications.
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Acknowledgments
JAG acknowledges funding from the Natural Environment Research Council (NE/C507902/1). KZ acknowledges funding from NHGRI (R01 HG004876). JDN acknowledges funding from a Discovery Grant from the National Sciences and Engineering Research Council of Canada (NSERC).
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Gilbert, J.A., Zhang, K., Neufeld, J.D. (2010). Multiple Displacement Amplification. In: Timmis, K.N. (eds) Handbook of Hydrocarbon and Lipid Microbiology. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-540-77587-4_333
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DOI: https://doi.org/10.1007/978-3-540-77587-4_333
Publisher Name: Springer, Berlin, Heidelberg
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