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Titolo:
Quantification of bacterial subgroups in soil: comparison of DNA extracteddirectly from soil or from cells previously released by density gradient centrifugation
Autore:
Courtois, S; Frostegard, A; Goransson, P; Depret, G; Jeannin, P; Simonet, P;
Indirizzi:
Univ Lyon 1, Lab Ecol Microbienne, CNRS, UMR 5557, F-69622 Villeurbanne, France Univ Lyon 1 Villeurbanne France F-69622 57, F-69622 Villeurbanne, France Aventis Pharma, Ctr Rech Vitry Alfortville, F-94403 Vitry Sur Seine, France Aventis Pharma Vitry Sur Seine France F-94403 03 Vitry Sur Seine, France Lund Univ, Dept Microbial Ecol, SE-22362 Lund, Sweden Lund Univ Lund Sweden SE-22362 ept Microbial Ecol, SE-22362 Lund, Sweden Agr Univ Norway, Dept Chem & Biotechnol, N-1432 As, Norway Agr Univ Norway As Norway N-1432 pt Chem & Biotechnol, N-1432 As, Norway
Titolo Testata:
ENVIRONMENTAL MICROBIOLOGY
fascicolo: 7, volume: 3, anno: 2001,
pagine: 431 - 439
SICI:
1462-2912(200107)3:7<431:QOBSIS>2.0.ZU;2-4
Fonte:
ISI
Lingua:
ENG
Soggetto:
POLYMERASE CHAIN-REACTION; RIBOSOMAL-RNA SEQUENCES; MICROBIAL-CELLS; PCR; AMPLIFICATION; SEPARATION; SEDIMENTS; GENOME; STRAIN; IDENTIFICATION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
46
Recensione:
Indirizzi per estratti:
Indirizzo: Simonet, P Univ Lyon 1, Lab Ecol Microbienne, CNRS, UMR 5557, 43 Bd 11 Novembre 1918,F-69622 Villeurbanne, France Univ Lyon 1 43 Bd 11 Novembre 1918 Villeurbanne France F-69622
Citazione:
S. Courtois et al., "Quantification of bacterial subgroups in soil: comparison of DNA extracteddirectly from soil or from cells previously released by density gradient centrifugation", ENVIRON MIC, 3(7), 2001, pp. 431-439

Abstract

All molecular analyses of soil bacterial diversity are based on the extraction of a representative fraction of cellular DNA. Methods of DNA extraction for this purpose are divided into two categories: those in which cells are lysed within the soil (direct extraction) and those in which cells are first removed from soil (cell extraction) and then lysed. The purpose of thisstudy was to compare a method of direct extraction with a method in which cells were first separated from the soil matrix by Nycodenz gradient centrifugation in order to evaluate the effect of these different approaches on the analysis of the spectrum of diversity in a microbial community. We used a method based on polymerase chain reaction (PCR) amplification of a 16S rRNA gene fragment, followed by hybridization of the amplified fragments to aset of specific probes to assess the phylogenetic diversity of our samples. Control parameters, such as the relationship between amount of DNA template and amount of PCR product and the influence of competing DNA on PCR amplification, were first examined. Comparison between extraction methods showed that less DNA was extracted when cells were first separated from the soilmatrix (0.4 mug g(-1) dry weight soil versus 38-93 mug g(-1) obtained by in situ lysis methods). However, with the exception of the gamma -subclass of Proteobacteria, there was no significant difference in the spectrum of diversity resulting from the two extraction strategies.

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Documento generato il 02/04/20 alle ore 00:24:24