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Microbiome

, 1:4

First Online: 09 January 2013Received: 30 March 2012Accepted: 23 May 2012

Abstract

BackgroundThe use of culture-independent nucleic acid techniques, such as ribosomal RNA gene cloning library analysis, has unveiled the tremendous microbial diversity that exists in natural environments. In sharp contrast to this great achievement is the current difficulty in cultivating the majority of bacterial species or phylotypes revealed by molecular approaches. Although recent new technologies such as metagenomics and metatranscriptomics can provide more functionality information about the microbial communities, it is still important to develop the capacity to isolate and cultivate individual microbial species or strains in order to gain a better understanding of microbial physiology and to apply isolates for various biotechnological applications.

ResultsWe have developed a new system to cultivate bacteria in an array of droplets. The key component of the system is the microbe observation and cultivation array MOCA, which consists of a Petri dish that contains an array of droplets as cultivation chambers. MOCA exploits the dominance of surface tension in small amounts of liquid to spontaneously trap cells in well-defined droplets on hydrophilic patterns. During cultivation, the growth of the bacterial cells across the droplet array can be monitored using an automated microscope, which can produce a real-time record of the growth. When bacterial cells grow to a visible microcolony level in the system, they can be transferred using a micropipette for further cultivation or analysis.

ConclusionsMOCA is a flexible system that is easy to set up, and provides the sensitivity to monitor growth of single bacterial cells. It is a cost-efficient technical platform for bioassay screening and for cultivation and isolation of bacteria from natural environments.

KeywordsMicrobe observation and cultivation array MOCA Cultivation Growth Microbiota AbbreviationsMOCAMicrobe observation and cultivation array

DNADeoxyribonucleic acid

rRNARibosomal ribonucleic acid

SSU rRNASmall subunit ribosomal RNA

PCRPolymerase chain reaction

μPLATMicroscale plasma activated templating

PDMSPolydimethylsiloxane.

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Autor: Weimin Gao - Dena Navarroli - Jared Naimark - Weiwen Zhang - Shih-hui Chao - Deirdre R Meldrum

Fuente: https://link.springer.com/article/10.1186/2049-2618-1-4







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