Cited 4 time in webofscience Cited 4 time in scopus

Lysis of a Single Cyanobacterium for Whole Genome Amplification

Title
Lysis of a Single Cyanobacterium for Whole Genome Amplification
Authors
Hall, EW[Hall, Eric W.]Kim, S[Kim, Samuel]Appadoo, V[Appadoo, Visham]Zare, RN[Zare, Richard N.]
DGIST Authors
Kim, S[Kim, Samuel]
Issue Date
2013-09
Citation
Micromachines, 4(3), 321-332
Type
Article
Article Type
Article
Keywords
CellsCyanobacteriaCytologyGenesLysisMicro-FluidicsMultiple Displacement AmplificationsNatural EnvironmentsSingle-CellSingle-Cell AnalysisSingle-CellsSynechocystis Sp. Pcc 6803Whole Genome AmplificationWhole Genome Amplifications
ISSN
2072-666X
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). © 2013 by the authors; licensee MDPI, Basel, Switzerland.
URI
http://hdl.handle.net/20.500.11750/3207
DOI
10.3390/mi4030321
Publisher
MDPI AG
Files:
There are no files associated with this item.
Collection:
New BiologyETC1. Journal Articles


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