{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T02:18:01Z","timestamp":1792635481681,"version":"4.5.0"},"reference-count":26,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2012,11,3]],"date-time":"2012-11-03T00:00:00Z","timestamp":1351900800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecular Ecology Resources"],"published-print":{"date-parts":[[2013,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The ubiquity, high diversity and often\u2010cryptic manifestations of fungi and oomycetes frequently necessitate molecular tools for detecting and identifying them in the environment. In applications including <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> barcoding, pathogen detection from plant samples, and genotyping for population genetics and epidemiology, rapid and dependable <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> extraction methods scalable from one to hundreds of samples are desirable. We evaluated several rapid extraction methods (<jats:styled-content style=\"fixed-case\">N<\/jats:styled-content>a<jats:styled-content style=\"fixed-case\">OH<\/jats:styled-content>, Rapid one\u2010step extraction (<jats:styled-content style=\"fixed-case\">ROSE<\/jats:styled-content>), Chelex 100, proteinase <jats:styled-content style=\"fixed-case\">K<\/jats:styled-content>) for their ability to obtain <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> of quantity and quality suitable for the following applications: <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> amplification of the multicopy barcoding locus <jats:styled-content style=\"fixed-case\">ITS<\/jats:styled-content>1\/5.8<jats:styled-content style=\"fixed-case\">S<\/jats:styled-content>\/<jats:styled-content style=\"fixed-case\">ITS<\/jats:styled-content>2 from various fungal cultures and sporocarps; single\u2010copy microsatellite amplification from cultures of the phytopathogenic oomycete <jats:italic>Phytophthora ramorum<\/jats:italic>; probe\u2010based <jats:italic>P.\u00a0ramorum<\/jats:italic> detection from leaves. Several methods were effective for most of the applications, with <jats:styled-content style=\"fixed-case\">N<\/jats:styled-content>a<jats:styled-content style=\"fixed-case\">OH<\/jats:styled-content> extraction favored in terms of success rate, cost, speed and simplicity. Frozen dilutions of <jats:styled-content style=\"fixed-case\">ROSE<\/jats:styled-content> and <jats:styled-content style=\"fixed-case\">N<\/jats:styled-content>a<jats:styled-content style=\"fixed-case\">OH<\/jats:styled-content> extracts maintained <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> viability for over 32\u00a0months. <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> from rapid extractions performed poorly compared to <jats:styled-content style=\"fixed-case\">CTAB<\/jats:styled-content>\/phenol\u2010chloroform extracts for <jats:styled-content style=\"fixed-case\">T<\/jats:styled-content>aq<jats:styled-content style=\"fixed-case\">M<\/jats:styled-content>an diagnostics from tanoak leaves, suggesting that incomplete removal of <jats:styled-content style=\"fixed-case\">PCR<\/jats:styled-content> inhibitors is an issue for sensitive diagnostic procedures, especially from plants with recalcitrant leaf chemistry. Na<jats:styled-content style=\"fixed-case\">OH<\/jats:styled-content> extracts exhibited lower yield and size than <jats:styled-content style=\"fixed-case\">CTAB<\/jats:styled-content>\/phenol\u2010chloroform extracts; however, <jats:styled-content style=\"fixed-case\">N<\/jats:styled-content>a<jats:styled-content style=\"fixed-case\">OH<\/jats:styled-content> extraction facilitated obtaining clean sequence data from sporocarps contaminated by other fungi, perhaps due to dilution resulting from low <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> yield. We conclude that conventional extractions are often unnecessary for routine <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> sequencing or genotyping of fungi and oomycetes, and recommend simpler strategies where source materials and intended applications warrant such use.<\/jats:p>","DOI":"10.1111\/1755-0998.12031","type":"journal-article","created":{"date-parts":[[2012,11,3]],"date-time":"2012-11-03T06:46:20Z","timestamp":1351925180000},"page":"66-74","source":"Crossref","is-referenced-by-count":72,"title":["Back to basics: an evaluation of <scp>N<\/scp>a<scp>OH<\/scp> and alternative rapid <scp>DNA<\/scp> extraction protocols for <scp>DNA<\/scp> barcoding, genotyping, and disease diagnostics from fungal and oomycete samples"],"prefix":"10.1111","volume":"13","author":[{"given":"Todd W.","family":"Osmundson","sequence":"first","affiliation":[{"name":"Forest Pathology and Mycology Laboratory Department of Environmental Science Policy &amp; Management University of California 137 Mulford Hall Berkeley CA 94720 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Catherine A.","family":"Eyre","sequence":"additional","affiliation":[{"name":"Forest Pathology and Mycology Laboratory Department of Environmental Science Policy &amp; Management University of California 137 Mulford Hall Berkeley CA 94720 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katherine M.","family":"Hayden","sequence":"additional","affiliation":[{"name":"Forest Pathology and Mycology Laboratory Department of Environmental Science Policy &amp; Management University of California 137 Mulford Hall Berkeley CA 94720 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jaskirn","family":"Dhillon","sequence":"additional","affiliation":[{"name":"Forest Pathology and Mycology Laboratory Department of Environmental Science Policy &amp; Management University of California 137 Mulford Hall Berkeley CA 94720 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matteo M.","family":"Garbelotto","sequence":"additional","affiliation":[{"name":"Forest Pathology and Mycology Laboratory Department of Environmental Science Policy &amp; Management University of California 137 Mulford Hall Berkeley CA 94720 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2012,11,3]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1080\/13693780600564613"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1755-0998.2009.02825.x"},{"key":"e_1_2_6_4_1","volume-title":"Phytophthora Diseases Worldwide","author":"Erwin 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