GC Fractionation Enhances Microbial CommunityDiversity Assessment and Detection of Minority Populations ofBacteria by Denaturing Gradient GelElectrophoresis
ABSTRACT Effectively and accurately assessing total microbial community diversity is one of the primary challenges in modern microbial ecology. This is particularly true with regard to the detection and characterization of unculturable populations and those present only in low abundance. We report a...
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| Published in | Applied and environmental microbiology Vol. 70; no. 4; pp. 2263 - 2270 |
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| Main Authors | , , , |
| Format | Journal Article |
| Language | English |
| Published |
01.04.2004
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| Online Access | Get full text |
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| Summary: | ABSTRACT
Effectively
and accurately assessing total microbial community diversity is one of
the primary challenges in modern microbial ecology. This is
particularly true with regard to the detection and characterization of
unculturable populations and those present only in low abundance. We
report a novel strategy, GC fractionation combined with denaturing
gradient gel electrophoresis (GC-DGGE), which combines mechanistically
different community analysis approaches to enhance assessment of
microbial community diversity and detection of minority populations of
microbes. This approach employs GC fractionation as an initial step to
reduce the complexity of the community in each fraction. This reduced
complexity facilitates subsequent detection of diversity in individual
fractions. DGGE analysis of individual fractions revealed bands that
were undetected or only poorly represented when total bacterial
community DNA was analyzed. Also, directed cloning and sequencing of
individual bands from DGGE lanes corresponding to individual
G+C fractions allowed detection of numerous phylotypes that
were not recovered using a traditional random cloning and sequencing
approach. |
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| ISSN: | 0099-2240 1098-5336 |
| DOI: | 10.1128/AEM.70.4.2263-2270.2004 |