Rapid and Effective DNA Extraction Method with Bead Grinding for a Large Amount of Fungal DNA
Abstract:To identify a rapid method for extracting a large amount of DNA from fungi associated with food hygiene, extraction methods were compared using fungal pellets formed rapidly in liquid media. Combinations of physical and chemical methods or commercial kits were evaluated with 3 species of yeast, 10 species of ascomycetous molds, and 4 species of zygomycetous molds. Bead grinding was the physical method, followed by chemical methods involving sodium dodecyl sulfate (SDS), cetyl trimethyl ammonium bromide (CTAB), and benzyl chloride and two commercial kits. Quantity was calculated by UV absorbance at 260 nm, quality was determined by the ratio of UV absorbance at 260 and 280 nm, and gene amplifications and electrophoresis profiles of whole genomes were analyzed. Bead grinding with the SDS method was the most effective for DNA extraction for yeasts and ascomycetous molds, and bead grinding with the CTAB method was most effective with zygomycetous molds. For both groups of molds, bead grinding with the CTAB method was the best approach for DNA extraction. Because this combination also is relatively effective for yeasts, it can be used to extract a large amount of DNA from a wide range of fungi. The DNA extraction methods are useful for developing gene indexes to identify fungi with molecular techniques, such as DNA fingerprinting.
Document Type: Research Article
Affiliations: 1: Division of Microbiology, National Institute of Health Sciences, Kamiyoga 1-18-1, Setagaya-ku, Tokyo 158-8501, Japan 2: Veterinary Medical Science, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657, Japan 3: Food Research Laboratories, Mitsui Norin Co., Ltd., Miyabara 223-1, Fujieda-shi, Shizuoka 426-0133, Japan
Publication date: June 1, 2010
- IAFP Members with personal subscriptions to JFP Online: To access full-text JFP or JMFT articles, you must sign-in in the upper-right corner using your Ingenta sign-in details (your IAFP Member Login does not apply to this website). The Journal of Food Protection (JFP) is a refereed monthly publication. Each issue contains scientific research and authoritative review articles reporting on a variety of topics in food science pertaining to food safety and quality. The Journal is internationally recognized as the leading publication in the field of food microbiology with a readership exceeding 11,000 scientists from 70 countries. The Journal of Food Protection is indexed in Index Medicus, Current Contents, BIOSIS, PubMed, Medline, and many others.
Print and online subscriptions are available to IAFP Members and institutional subscribers. IAFP Members with a subscription to JFP Online will have access to all available JFP and JMFT content. Online visitors who are not IAFP Members or journal subscribers will be charged on a pay-per-view basis. Membership and subscription information is available at www.foodprotection.org.
- Information for Authors
- Submit a Paper
- Subscribe to this Title
- Membership Information
- Information for Advertisers
- ingentaconnect is not responsible for the content or availability of external websites