Rapid method for direct extraction of DNA from soil and sediments.
Yu-Li Tsai,Betty H. Olson +1 more
TLDR
A rapid method for the direct extraction of DNA from soil and sediments was developed, which allows for the analysis of small samples and the processing of many samples in a relatively short (7 h) period.Abstract:
A rapid method for the direct extraction of DNA from soil and sediments was developed. The indigenous microorganisms in the soil and sediments were lysed by using lysozyme and a freeze-thaw procedure. The lysate was extracted with sodium dodecyl sulfate and phenol-chloroform. In addition to a high recovery efficiency (greater than 90%), the yields of DNA were high (38 and 12 micrograms/g [wet weight] from sediments and soil, respectively). This method generated minimal shearing of the extracted DNA. The crude DNA could be further purified with an Elutip-d column if necessary. An additional advantage of this method is that only 1 g of sample is required, which allows for the analysis of small samples and the processing of many samples in a relatively short (7 h) period.read more
Citations
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Journal ArticleDOI
Determination of microbial diversity in environmental samples: pitfalls of PCR‐based rRNA analysis
TL;DR: Specific aspects of sample collection, cell lysis, nucleic acid extraction, PCR amplification, separation of amplified DNA, application of nucleic probes and data analysis are covered.
Journal ArticleDOI
Bacterial gene transfer by natural genetic transformation in the environment.
TL;DR: The current understanding of the biology of transformation is summarized to provide the platform on which aspects of bacterial transformation in water, soil, and sediments and the habitat of pathogens are discussed.
Journal ArticleDOI
Evaluation and Optimization of DNA Extraction and Purification Procedures for Soil and Sediment Samples
TL;DR: It is indicated that for these types of samples, optimum DNA recovery requires brief, low-speed bead mill homogenization in the presence of a phosphate-buffered SDS-chloroform mixture, followed by Sephadex G-200 column purification.
Journal ArticleDOI
Microbial Evolution, Diversity, and Ecology: A Decade of Ribosomal RNA Analysis of Uncultivated Microorganisms
TL;DR: This review provides an outline of the main methods used in molecular microbial ecology, and their limitations, with reference to morphologically distinctive, uncultivated bacteria; an important biotechnological process (wastewater treatment); and symbiotic relationships between Bacteria, Archaea and Eukarya.
Journal ArticleDOI
Occurrence and Diversity of Tetracycline Resistance Genes in Lagoons and Groundwater Underlying Two Swine Production Facilities
TL;DR: It is demonstrated that tet genes occur in the environment as a direct result of agriculture and suggested that groundwater may be a potential source of antibiotic resistance in the food chain.
References
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Book
Molecular Cloning: A Laboratory Manual
TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
UseofNuclepore Filters forCounting Bacteria by Fluorescence Microscopy
TL;DR: Polycarbonate Nuclepore filters are better than cellulose filters for the direct counting of bacteria because they have uniform pore size and a flat surface that retains all of the bacteria on top of the filter.
Journal ArticleDOI
Use of nuclepore filters for counting bacteria by fluorescence microscopy.
TL;DR: Polycarbonate Nuclepore filters are better than cellulose filters for the direct counting of bacteria because they have uniform pore size and a flat surface that retains all of the bacteria on top of the filter.
Book
Introduction to soil microbiology
TL;DR: In this paper, the biological processes that take place in the soil and their importance to soil fertility, plant growth, and environmental quality are investigated from both descriptive and functional viewpoints, including microbial ecology, the carbon and nitrogen cycles, mineral transformation, and ecological interrelationships.
Journal ArticleDOI
Survival strategies of bacteria in the natural environment.
D B Roszak,Rita R. Colwell +1 more
TL;DR: The heterotrophic plate count has come under increasing criticism because it is inefficient, at best, for enumerating viable bacteria present in marine and estuarine systems.