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Lysis of a Single Cyanobacterium for Whole Genome Amplification
Department of Chemistry, Stanford University, Stanford, CA 94305, USA
Department of New Biology, DGIST, Daegu 711-873, Korea
Present Address: SRI International Inc., Menlo Park, CA 94025, USA
Present Address: Department of Chemistry, University of Wisconsin-Madison, Madison, WI 53706, USA
These authors contributed equally to this work.
* Author to whom correspondence should be addressed.
Received: 7 June 2013; in revised form: 8 July 2013 / Accepted: 12 August 2013 / Published: 21 August 2013
Abstract: Bacterial species from natural environments, exhibiting a great degree of genetic diversity that has yet to be characterized, pose a specific challenge to whole genome amplification (WGA) from single cells. A major challenge is establishing an effective, compatible, and controlled lysis protocol. We present a novel lysis protocol that can be used to extract genomic information from a single cyanobacterium of Synechocystis sp. PCC 6803 known to have multilayer cell wall structures that resist conventional lysis methods. Simple but effective strategies for releasing genomic DNA from captured cells while retaining cellular identities for single-cell analysis are presented. Successful sequencing of genetic elements from single-cell amplicons prepared by multiple displacement amplification (MDA) is demonstrated for selected genes (15 loci nearly equally spaced throughout the main chromosome).
Keywords: cyanobacteria; lysis; whole genome amplification; single cell; microfluidics
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MDPI and ACS Style
Hall, E.W.; Kim, S.; Appadoo, V.; Zare, R.N. Lysis of a Single Cyanobacterium for Whole Genome Amplification. Micromachines 2013, 4, 321-332.
Hall EW, Kim S, Appadoo V, Zare RN. Lysis of a Single Cyanobacterium for Whole Genome Amplification. Micromachines. 2013; 4(3):321-332.
Hall, Eric W.; Kim, Samuel; Appadoo, Visham; Zare, Richard N. 2013. "Lysis of a Single Cyanobacterium for Whole Genome Amplification." Micromachines 4, no. 3: 321-332.