4.5 Article

Evaluation of the denaturing gradient gel electrophoresis-apparatus as a parameter influencing soil microbial community fingerprinting

期刊

WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
卷 26, 期 9, 页码 1721-1726

出版社

SPRINGER
DOI: 10.1007/s11274-010-0349-z

关键词

Cattle impact; DGGE; DGGE interfering factors; Soil microbial community fingerprinting

资金

  1. Ente Cassa di Risparmio di Firenze
  2. Ministry of Education of the Czech Republic [LC06066]
  3. Grant Agency of the Academy of Sciences of the Czech Republic [IAA600660605]

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We compared two denaturing gradient gel electrophoresis (DGGE) systems-DCode (Biorad, Hercules, CA, USA) and PhorU (Ingeny, Leiden, NL), performing community level 16S and 18S rRNA gene fragment-PCR-DGGE with total DNA extracted from upland pasture soil used for outdoor cattle husbandry. The methodological evaluation of the DGGE apparatus as parameter influencing DGGE fingerprinting, based on cluster analysis of soil bacterial and fungal community fingerprints, was made in terms of the resulting information about microbial community structures and their response to different degrees of cattle impact. Although the comparative DGGE analysis with different DGGE systems provided similar clustering of microbial community structures in correlation with the degree of cattle impact, our results suggest the DGGE system to be a factor influencing DGGE analysis. To our knowledge this is the first attempt to investigate the hypothetical impact of the DGGE system due to different technical characteristics, recommending the use of one and the same DGGE apparatus throughout an experiment, if the monitoring of microbial community structures requires multiple gel-to-gel analysis.

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