A small-scale procedure for extracting nucleic acids from woody plants infected with various phytopathogens for PCR assay
The complexity of most nucleic acid extraction procedures limits the number of samples that can be easily processed for analysis by polymerase chain reaction (PCR). A simple, small-scale procedure was developed which can be carried out entirely in 1.5-ml microfuge tubes whereby the container and con...
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Veröffentlicht in: | Journal of virological methods 1998-03, Vol.71 (1), p.45-50 |
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description | The complexity of most nucleic acid extraction procedures limits the number of samples that can be easily processed for analysis by polymerase chain reaction (PCR). A simple, small-scale procedure was developed which can be carried out entirely in 1.5-ml microfuge tubes whereby the container and contents are frozen with liquid nitrogen, tissue is pulverized, and targeted nucleic acids are extracted. DNA of bacterial and phytoplasmal plant pathogens was extracted in hot CTAB buffer followed by chloroform clarification. Following centrifugation, the DNA in the aqueous fraction was precipitated with isopropanol and resuspended in water. RNA originating from viruses and viroids was extracted from triturated tissue using STE buffer and phenol. The nucleic acid fraction was purified using CF-11 cellulose. All purified preparations were used as PCR or RT-PCR templates to detect DNA or RNA, respectively. These procedures were used to detect
Xylella fastidiosa, peach yellow leaf roll phytoplasma, sour cherry green ring mottle virus, and peach latent mosaic viroid by agarose gel electrophoresis. |
doi_str_mv | 10.1016/S0166-0934(97)00190-0 |
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Xylella fastidiosa, peach yellow leaf roll phytoplasma, sour cherry green ring mottle virus, and peach latent mosaic viroid by agarose gel electrophoresis.</description><identifier>ISSN: 0166-0934</identifier><identifier>EISSN: 1879-0984</identifier><identifier>DOI: 10.1016/S0166-0934(97)00190-0</identifier><identifier>PMID: 9628220</identifier><identifier>CODEN: JVMEDH</identifier><language>eng</language><publisher>London: Elsevier B.V</publisher><subject>ACIDE NUCLEIQUE ; ACIDOS NUCLEICOS ; AGENT PATHOGENE ; Bacterium ; Biological and medical sciences ; DNA, Bacterial - genetics ; DNA, Bacterial - isolation & purification ; Electrophoresis, Agar Gel ; Fundamental and applied biological sciences. Psychology ; Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control ; METHODE ; METHODS ; METODOS ; Microbiology ; Nucleic acid extraction ; NUCLEIC ACIDS ; ORGANISMOS PATOGENOS ; PATHOGENS ; PCR ; Phytopathology. Animal pests. Plant and forest protection ; Phytoplasma ; Plant viruses and viroids ; PLANTAS LENOSAS ; PLANTE LIGNEUSE ; Plants - microbiology ; Plants - virology ; Polymerase Chain Reaction - methods ; RNA, Viral - genetics ; RNA, Viral - isolation & purification ; Rosales - microbiology ; Rosales - virology ; Techniques used in virology ; Viroid ; Virology ; Virus ; WOODY PLANTS</subject><ispartof>Journal of virological methods, 1998-03, Vol.71 (1), p.45-50</ispartof><rights>1998 Elsevier Science B.V.</rights><rights>1998 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c442t-659e45eb4ad94d6f056107bcaa1b142c2c97eee6a3f3b2a3258b452fa6a880ea3</citedby><cites>FETCH-LOGICAL-c442t-659e45eb4ad94d6f056107bcaa1b142c2c97eee6a3f3b2a3258b452fa6a880ea3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/S0166-0934(97)00190-0$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2274908$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9628220$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zhang, Yun-ping</creatorcontrib><creatorcontrib>Uyemoto, J.K</creatorcontrib><creatorcontrib>Kirkpatrick, B.C</creatorcontrib><title>A small-scale procedure for extracting nucleic acids from woody plants infected with various phytopathogens for PCR assay</title><title>Journal of virological methods</title><addtitle>J Virol Methods</addtitle><description>The complexity of most nucleic acid extraction procedures limits the number of samples that can be easily processed for analysis by polymerase chain reaction (PCR). A simple, small-scale procedure was developed which can be carried out entirely in 1.5-ml microfuge tubes whereby the container and contents are frozen with liquid nitrogen, tissue is pulverized, and targeted nucleic acids are extracted. DNA of bacterial and phytoplasmal plant pathogens was extracted in hot CTAB buffer followed by chloroform clarification. Following centrifugation, the DNA in the aqueous fraction was precipitated with isopropanol and resuspended in water. RNA originating from viruses and viroids was extracted from triturated tissue using STE buffer and phenol. The nucleic acid fraction was purified using CF-11 cellulose. All purified preparations were used as PCR or RT-PCR templates to detect DNA or RNA, respectively. These procedures were used to detect
Xylella fastidiosa, peach yellow leaf roll phytoplasma, sour cherry green ring mottle virus, and peach latent mosaic viroid by agarose gel electrophoresis.</description><subject>ACIDE NUCLEIQUE</subject><subject>ACIDOS NUCLEICOS</subject><subject>AGENT PATHOGENE</subject><subject>Bacterium</subject><subject>Biological and medical sciences</subject><subject>DNA, Bacterial - genetics</subject><subject>DNA, Bacterial - isolation & purification</subject><subject>Electrophoresis, Agar Gel</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control</subject><subject>METHODE</subject><subject>METHODS</subject><subject>METODOS</subject><subject>Microbiology</subject><subject>Nucleic acid extraction</subject><subject>NUCLEIC ACIDS</subject><subject>ORGANISMOS PATOGENOS</subject><subject>PATHOGENS</subject><subject>PCR</subject><subject>Phytopathology. Animal pests. Plant and forest protection</subject><subject>Phytoplasma</subject><subject>Plant viruses and viroids</subject><subject>PLANTAS LENOSAS</subject><subject>PLANTE LIGNEUSE</subject><subject>Plants - microbiology</subject><subject>Plants - virology</subject><subject>Polymerase Chain Reaction - methods</subject><subject>RNA, Viral - genetics</subject><subject>RNA, Viral - isolation & purification</subject><subject>Rosales - microbiology</subject><subject>Rosales - virology</subject><subject>Techniques used in virology</subject><subject>Viroid</subject><subject>Virology</subject><subject>Virus</subject><subject>WOODY PLANTS</subject><issn>0166-0934</issn><issn>1879-0984</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1998</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkVtv1DAQhSMEKkvhJ1TyA0LlIeBL4thPqFpBi7QCxOXZmjiTXaMkDnbSkn-P96LlsS-25HM8c2a-LLti9B2jTL7_kQ6ZUy2Ka129pZRpmtMn2YqpSqdnVTzNVmfL8-xFjL8ppWUlxEV2oSVXnNNVttyQ2EPX5dFCh2QM3mIzByStDwT_TgHs5IYtGWbbobMErGsiaYPvyYP3zULGDoYpEje0aCdsyIObduQegvNzJONumfwI085vcYiHmt_W3wnECMvL7FkLXcRXp_sy-_Xp48_1Xb75evt5fbPJbVHwKZelxqLEuoBGF41saSkZrWoLwGpWcMutrhBRgmhFzUHwUtVFyVuQoBRFEJfZm2PdNNufGeNkehctdik3poym0lpwJeijRiY5laUUyVgejTb4GAO2Zgyuh7AYRs2ejTmwMfvFG12ZAxuzb3B1ajDXPTbnXycYSX990mFPow0wWBfPNs6rQlP1v0wL3sA2JMuXDdNaUcqV0kn_cNQxbfXeYTDROhwSVxcSI9N490jQf9EStbA</recordid><startdate>19980301</startdate><enddate>19980301</enddate><creator>Zhang, Yun-ping</creator><creator>Uyemoto, J.K</creator><creator>Kirkpatrick, B.C</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7T7</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>19980301</creationdate><title>A small-scale procedure for extracting nucleic acids from woody plants infected with various phytopathogens for PCR assay</title><author>Zhang, Yun-ping ; Uyemoto, J.K ; Kirkpatrick, B.C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c442t-659e45eb4ad94d6f056107bcaa1b142c2c97eee6a3f3b2a3258b452fa6a880ea3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1998</creationdate><topic>ACIDE NUCLEIQUE</topic><topic>ACIDOS NUCLEICOS</topic><topic>AGENT PATHOGENE</topic><topic>Bacterium</topic><topic>Biological and medical sciences</topic><topic>DNA, Bacterial - genetics</topic><topic>DNA, Bacterial - isolation & purification</topic><topic>Electrophoresis, Agar Gel</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control</topic><topic>METHODE</topic><topic>METHODS</topic><topic>METODOS</topic><topic>Microbiology</topic><topic>Nucleic acid extraction</topic><topic>NUCLEIC ACIDS</topic><topic>ORGANISMOS PATOGENOS</topic><topic>PATHOGENS</topic><topic>PCR</topic><topic>Phytopathology. Animal pests. Plant and forest protection</topic><topic>Phytoplasma</topic><topic>Plant viruses and viroids</topic><topic>PLANTAS LENOSAS</topic><topic>PLANTE LIGNEUSE</topic><topic>Plants - microbiology</topic><topic>Plants - virology</topic><topic>Polymerase Chain Reaction - methods</topic><topic>RNA, Viral - genetics</topic><topic>RNA, Viral - isolation & purification</topic><topic>Rosales - microbiology</topic><topic>Rosales - virology</topic><topic>Techniques used in virology</topic><topic>Viroid</topic><topic>Virology</topic><topic>Virus</topic><topic>WOODY PLANTS</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhang, Yun-ping</creatorcontrib><creatorcontrib>Uyemoto, J.K</creatorcontrib><creatorcontrib>Kirkpatrick, B.C</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of virological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhang, Yun-ping</au><au>Uyemoto, J.K</au><au>Kirkpatrick, B.C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A small-scale procedure for extracting nucleic acids from woody plants infected with various phytopathogens for PCR assay</atitle><jtitle>Journal of virological methods</jtitle><addtitle>J Virol Methods</addtitle><date>1998-03-01</date><risdate>1998</risdate><volume>71</volume><issue>1</issue><spage>45</spage><epage>50</epage><pages>45-50</pages><issn>0166-0934</issn><eissn>1879-0984</eissn><coden>JVMEDH</coden><abstract>The complexity of most nucleic acid extraction procedures limits the number of samples that can be easily processed for analysis by polymerase chain reaction (PCR). A simple, small-scale procedure was developed which can be carried out entirely in 1.5-ml microfuge tubes whereby the container and contents are frozen with liquid nitrogen, tissue is pulverized, and targeted nucleic acids are extracted. DNA of bacterial and phytoplasmal plant pathogens was extracted in hot CTAB buffer followed by chloroform clarification. Following centrifugation, the DNA in the aqueous fraction was precipitated with isopropanol and resuspended in water. RNA originating from viruses and viroids was extracted from triturated tissue using STE buffer and phenol. The nucleic acid fraction was purified using CF-11 cellulose. All purified preparations were used as PCR or RT-PCR templates to detect DNA or RNA, respectively. These procedures were used to detect
Xylella fastidiosa, peach yellow leaf roll phytoplasma, sour cherry green ring mottle virus, and peach latent mosaic viroid by agarose gel electrophoresis.</abstract><cop>London</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>9628220</pmid><doi>10.1016/S0166-0934(97)00190-0</doi><tpages>6</tpages></addata></record> |
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subjects | ACIDE NUCLEIQUE ACIDOS NUCLEICOS AGENT PATHOGENE Bacterium Biological and medical sciences DNA, Bacterial - genetics DNA, Bacterial - isolation & purification Electrophoresis, Agar Gel Fundamental and applied biological sciences. Psychology Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control METHODE METHODS METODOS Microbiology Nucleic acid extraction NUCLEIC ACIDS ORGANISMOS PATOGENOS PATHOGENS PCR Phytopathology. Animal pests. Plant and forest protection Phytoplasma Plant viruses and viroids PLANTAS LENOSAS PLANTE LIGNEUSE Plants - microbiology Plants - virology Polymerase Chain Reaction - methods RNA, Viral - genetics RNA, Viral - isolation & purification Rosales - microbiology Rosales - virology Techniques used in virology Viroid Virology Virus WOODY PLANTS |
title | A small-scale procedure for extracting nucleic acids from woody plants infected with various phytopathogens for PCR assay |
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