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Micromachines 2013, 4(3), 321-332; doi:10.3390/mi4030321
Article

Lysis of a Single Cyanobacterium for Whole Genome Amplification

1,§,†
,
1,2,†
,
1,‡
 and
1,*
Received: 7 June 2013 / Revised: 8 July 2013 / Accepted: 12 August 2013 / Published: 21 August 2013
(This article belongs to the Special Issue Micro/Nanofluidic Devices for Single Cell Analysis)
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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 cyanobacteria; lysis; whole genome amplification; single cell; microfluidics
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Hall, E.W.; Kim, S.; Appadoo, V.; Zare, R.N. Lysis of a Single Cyanobacterium for Whole Genome Amplification. Micromachines 2013, 4, 321-332.

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