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Titolo:
Comparison of different methods for the isolation and purification of total community DNA from soil
Autore:
Krsek, M; Wellington, EMH;
Indirizzi:
Univ Warwick, Dept Biol Sci, Coventry CV4 7AL, W Midlands, England Univ Warwick Coventry W Midlands England CV4 7AL 7AL, W Midlands, England
Titolo Testata:
JOURNAL OF MICROBIOLOGICAL METHODS
fascicolo: 1, volume: 39, anno: 1999,
pagine: 1 - 16
SICI:
0167-7012(199912)39:1<1:CODMFT>2.0.ZU;2-N
Fonte:
ISI
Lingua:
ENG
Soggetto:
GRADIENT GEL-ELECTROPHORESIS; POLYMERASE CHAIN-REACTION; 16S RIBOSOMAL-RNA; ENVIRONMENTAL-SAMPLES; MICROBIAL DNA; PLASMID DNA; EXTRACTION; BACTERIA; PCR; DIVERSITY;
Keywords:
DNA extraction; microbial community analysis; soil microbiology; total community DNA;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
38
Recensione:
Indirizzi per estratti:
Indirizzo: Wellington, EMH Univ Warwick, Dept Biol Sci, Coventry CV4 7AL, W Midlands,England Univ Warwick Coventry W Midlands England CV4 7AL England
Citazione:
M. Krsek e E.M.H. Wellington, "Comparison of different methods for the isolation and purification of total community DNA from soil", J MICROB M, 39(1), 1999, pp. 1-16

Abstract

The efficiency and reproducibility of DNA extraction from soil was tested for variations in lyric and purification treatments and their effect on yield and purity of DNA. The extraction yield was improved by increasing the concentration of EDTA or monovalent ions in isolation buffers,, by the introduction of mechanical lysis treatments, and by the use of ethanol precipitation in place of PEG precipitation. Purity was improved using buffers with decreasing concentration of EDTA or by reducing the ionic strength of the buffer, and by all mechanical treatments. No lyric treatment was efficient on its own, the highest purity was achieved using Crombach buffer and a combination of bead-beating with lysozyme and SDS lysis followed by potassium acetate and PEG precipitation, phenol/chloroform purification, isopropanol precipitation, and spermine-HCl precipitation. Sonication sheared the DNA more than bead-heating. Lysozyme and SDS lysis without any mechanical treatments allowed isolation of larger fragments (40-90 kb). Denaturing gradient gel electrophoresis analysis of DNA isolated using a range of lyric treatments revealed alterations in band patterns which might reflect differences inthe efficiency of lytic treatments. (C) 1999 Elsevier Science B.V. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 08/07/20 alle ore 01:01:14