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Title 

Development of microbial genome-probing microarrays using digital multiple displacement amplification of uncultivated microbial single cells

 

미배양 단일 세포의 디지털 다중증폭법을 통한 미배양 미생물 모니터링용 GPM 개발

Authors 

Ho Won ChangY SungKyoung-Ho KimYoung Do NamSeong Woon RohMin-Su KimC O JeonJin-Woo Bae

Publisher 

American Chemical Society

Issue Date 

2008

Citation 

Environmental Science & Technology, vol. 42, no. 16, pp. 6058-6064

Keywords 

digital arithmeticmicroarraysbacterial cellsbacterial genomesgenome-probing microarraysmicrobial detectionmicrobial diversitymicrobial populationsmultiple displacement amplificationgenome

Abstract 

A crucial problem in the use of previously developed genome-probing microarrays (GPM) has been the inability to use uncultivated bacterial genomes to take advantage of the high sensitivity and specificity of GPM in microbial detection and monitoring. We show here a method, digital multiple displacement amplification (MDA), to amplify and analyze various genomes obtained from single uncultivated bacterial cells. We used 15 genomes from key microbes involved in dichloromethane (DCM)-dechlorinating enrichment as microarray probes to uncover the bacterial population dynamics of samples without PCR amplification. Genomic DNA amplified from single cells originating from uncultured bacteria with 80.3-99.4% similarity to 16S rRNA genes of cultivated bacteria. The digital MDA-GPM method successfully monitored the dynamics of DCM-dechlorinating communities from different phases of enrichment status. Without a priori knowledge of microbial diversity, the digital MDA-GPM method could be designed to monitor most microbial populations in a given environmental sample.

ISSN 

0013-936X

Link 

http://dx.doi.org/10.1021/es8006029

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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