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@ARTICLE{Ning:7048,
author = {Ning, J. and Liebich, J. and Kästner, M. and Zhou, J. Z.
and Schäffer, A. and Burauel, P.},
title = {{D}ifferent influences of {DNA} purity indices and quantity
on {PCR}-based {DGGE} and functional gene microarray in soil
microbial community study},
journal = {Applied Microbiology and Biotechnology},
volume = {82},
issn = {0175-7598},
address = {Berlin},
publisher = {Springer},
reportid = {PreJuSER-7048},
pages = {983 - 993},
year = {2009},
note = {The manuscript is a joint approach within the working group
"Biodiversity" of the Helmholtz Association, Germany. The
authors would like to thank Eva Mareike Seeger, Mareike
Braeckevelt, and Heidrun Paschke in the group of the
Department of Bioremediation (UFZ, Germany) for the help
during the sampling at the SAFIRA plant in Bitterfeld and
the group in the Institute for Environmental Genomics
(Oklahoma, USA) for providing the microarray chips, as well
as Dr. Naixin Li for the valuable discussion. The
construction of microarray chips and its functional gene
array predecessors is supported by the Environmental Stress
Pathway Project (ESPP) of the Virtual Institute for
Microbial Stress and Survival (http://vimss. lbl. gov)
supported by the U. S. Department of Energy, Office of
Science, Office of Biological and Environmental Research,
Genomics: GTL Program, through contract DE- AC0205CH11231
with Lawrence Berkeley National Laboratory.},
abstract = {Based on the comparative study of the DNA extracts from two
soil samples obtained by three commercial DNA extraction
kits, we evaluated the influence of the DNA quantity and
purity indices (the absorbance ratios A260/280 and A260/230,
as well as the absorbance value A320 indicating the amount
of humic substances) on polymerase chain reaction
(PCR)-based denaturing gradient gel electrophoresis (DGGE)
and a functional gene microarray used in the study of
microbial communities. Numbers and intensities of the DGGE
bands are more affected by the A260/280 and A320 values than
by the ratio A260/230 and conditionally affected by the DNA
yield. Moreover, we demonstrated that the DGGE band pattern
was also affected by the preferential extraction due to
chemical agents applied in the extraction. Unlike DGGE,
microarray is more affected by the A260/230 and A320 values.
Until now, the successful PCR performance is the mostly used
criterion for soil DNA purity. However, since PCR was more
influenced by the A260/280 ratio than by A260/230, it is not
accurate enough any more for microbial community assessed by
non-PCR-based methods such as microarray. This study
provides some useful hints on how to choose effective DNA
extraction method for the subsequent assessment of microbial
community.},
keywords = {Bacteriological Techniques: methods / Biodiversity / DNA,
Bacterial: isolation $\&$ purification / Electrophoresis,
Gel, Two-Dimensional: methods / Humic Substances: analysis /
Oligonucleotide Array Sequence Analysis: methods /
Polymerase Chain Reaction: methods / Reagent Kits,
Diagnostic: standards / Reproducibility of Results / Soil
Microbiology / DNA, Bacterial (NLM Chemicals) / Humic
Substances (NLM Chemicals) / Reagent Kits, Diagnostic (NLM
Chemicals) / J (WoSType)},
cin = {ICG-4},
ddc = {570},
cid = {I:(DE-Juel1)VDB793},
pnm = {Terrestrische Umwelt},
pid = {G:(DE-Juel1)FUEK407},
shelfmark = {Biotechnology $\&$ Applied Microbiology},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:19247649},
UT = {WOS:000264460500020},
doi = {10.1007/s00253-009-1912-0},
url = {https://juser.fz-juelich.de/record/7048},
}