Literature DB >> 11295193

A strategy for optimizing quality and quantity of DNA extracted from soil.

H Bürgmann1, M Pesaro, F Widmer, J Zeyer.   

Abstract

The efficiency of a bead beating method was studied in detail with regard to a variety of factors including beating time and speed, volume and temperature of the buffer, as well as amount and type of beads employed. The results presented here reveal that all of these parameters have a significant effect on yield and quality of DNA extracted from soils. Precise adjustment of extraction conditions allows for significantly higher yields of high quality DNA from soils than previously reported. We further evaluated the effect of the extraction conditions on the apparent soil microbial community structures, as observed by polymerase chain reaction (PCR) and RFLP. Differences in the fingerprints of DNA extracted under different conditions suggest that results could be biased when using gentle extraction procedures. Based on multiple subsequent extractions using very harsh extraction conditions, we propose a protocol for the quantification of the total DNA content in soils. Extractions from six soils of different texture and chemical characteristics with selected bead beating protocols revealed that the quality (fragment size and purity) of the extracted DNA was generally very good, but also depended on the soil characteristics. While a single, general protocol for optimal DNA recovery from all soils cannot be given, this study provides detailed guidelines on how to optimize the general method to obtain optimal DNA from individual soils.

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Year:  2001        PMID: 11295193     DOI: 10.1016/s0167-7012(01)00213-5

Source DB:  PubMed          Journal:  J Microbiol Methods        ISSN: 0167-7012            Impact factor:   2.363


  61 in total

1.  Novel kingdom-level eukaryotic diversity in anoxic environments.

Authors:  Scott C Dawson; Norman R Pace
Journal:  Proc Natl Acad Sci U S A       Date:  2002-06-11       Impact factor: 11.205

2.  A census of rRNA genes and linked genomic sequences within a soil metagenomic library.

Authors:  Mark R Liles; Brian F Manske; Scott B Bintrim; Jo Handelsman; Robert M Goodman
Journal:  Appl Environ Microbiol       Date:  2003-05       Impact factor: 4.792

3.  New molecular screening tools for analysis of free-living diazotrophs in soil.

Authors:  Helmut Bürgmann; Franco Widmer; William Von Sigler; Josef Zeyer
Journal:  Appl Environ Microbiol       Date:  2004-01       Impact factor: 4.792

4.  Impact of soil drying-rewetting stress on microbial communities and activities and on degradation of two crop protection products.

Authors:  Manuel Pesaro; Gilles Nicollier; Josef Zeyer; Franco Widmer
Journal:  Appl Environ Microbiol       Date:  2004-05       Impact factor: 4.792

5.  Abundance, diversity and activity of sulfate-reducing prokaryotes in heavy metal-contaminated sediment from a salt marsh in the Medway Estuary (UK).

Authors:  Laurent Quillet; Ludovic Besaury; Milka Popova; Sandrine Paissé; Julien Deloffre; Baghdad Ouddane
Journal:  Mar Biotechnol (NY)       Date:  2011-11-30       Impact factor: 3.619

6.  A Simple Method for the Efficient Isolation of Genomic DNA from Lactobacilli Isolated from Traditional Indian Fermented Milk (dahi).

Authors:  Sachinandan De; Gurpreet Kaur; Amit Roy; Gaurav Dogra; Ramakant Kaushik; Paras Yadav; Rameshwar Singh; Tirtha Kumar Datta; Surender Lal Goswami
Journal:  Indian J Microbiol       Date:  2011-01-30       Impact factor: 2.461

7.  Practical method for extraction of PCR-quality DNA from environmental soil samples.

Authors:  Kelly A Fitzpatrick; Gilbert J Kersh; Robert F Massung
Journal:  Appl Environ Microbiol       Date:  2010-04-30       Impact factor: 4.792

8.  Effect of DNA extraction method on the apparent microbial diversity of soil.

Authors:  Ozgül Inceoglu; Eelco F Hoogwout; Patrick Hill; Jan Dirk van Elsas
Journal:  Appl Environ Microbiol       Date:  2010-03-26       Impact factor: 4.792

9.  Impacts of inter- and intralaboratory variations on the reproducibility of microbial community analyses.

Authors:  Yao Pan; Levente Bodrossy; Peter Frenzel; Anne-Grethe Hestnes; Sascha Krause; Claudia Lüke; Marion Meima-Franke; Henri Siljanen; Mette M Svenning; Paul L E Bodelier
Journal:  Appl Environ Microbiol       Date:  2010-09-24       Impact factor: 4.792

10.  Culture-independent analysis of the soil bacterial assemblage at the Great Salt Plains of Oklahoma.

Authors:  Ingrid R Caton; Mark A Schneegurt
Journal:  J Basic Microbiol       Date:  2011-09-23       Impact factor: 2.281

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