Applications of magnetic beads with covalently attached oligonucleotides in hybridization: Isolation and detection of specific measles virus mRNA from a crude cell lysate
Identifieur interne : 000E81 ( Istex/Corpus ); précédent : 000E80; suivant : 000E82Applications of magnetic beads with covalently attached oligonucleotides in hybridization: Isolation and detection of specific measles virus mRNA from a crude cell lysate
Auteurs : Catrine Albretsen ; Karl-Henning Kalland ; Bj Rn-Ivar Haukanes ; Leiv-Sigve H Varstein ; Kjell KleppeSource :
- Analytical Biochemistry [ 0003-2697 ] ; 1990.
English descriptors
- Teeft :
- Albretsen, Ammonia, Ammonolysis, Aqueous ammonia, Autoradiography, Bead, Bead surface, Beads, Blot analysis, Blot filter, Cdna, Cell lysate, Cellulose nitrate filter hybridization, Cellulose nitrate filters, Crude cell lysate, Detection limit, Different amounts, Different linker lengths, Eluate, External magnet, First wash, Guanidine thiocyanate, Guscn, Herring sperm, Hmrna, Hybridization, Hybridization solution, Linker, Linker length, Liquid scintillation, Lysate, Magnetic beads, Main chain, Measles, Measles cells, Measles virus, Methoxy phosphoramidites, Mrna, Negative control, Nonspecific binding, Nucleic, Nucleic acids, Nucleoside, Oligonucleotide, Oligonucleotide synthesis, Oligonucleotides, Plasmid, Prehybridization, Radiolabeled oligonucleotide, Room temperature, Sandwich hybridization, Scintillation, Solid support, Synthesizer, Target plasmid, Uninfected cells, Urethane group.
Abstract
Abstract: A novel, rapid, one-step isolation procedure utilizing oligonucleotides end-attached to magnetic beads (Dynabeads) has been developed. The beads and their captured target nucleic acids were separated after 2 h from the hybridization solution using an external magnet. This procedure was used to isolate measles virus hemagglutinin (H) mRNA from infected cells dissolved in 5 m guanidine thiocyanate. The yield of the specific, undegraded H-mRNA was found to be near the estimated total amount of H-mRNA present in the cells. The magnetic beads were also used for detection of measles virus H-mRNA in a crude cell lysate by sandwich hybridization. The experimental detection limit was 630 amol H-mRNA.
Url:
DOI: 10.1016/0003-2697(90)90041-7
Links to Exploration step
ISTEX:F55E88B7AFF080D43CBDABCCCA291DEFEB816599Le document en format XML
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Sigve" sort="H Varstein, Leiv Sigve" uniqKey="H Varstein L" first="Leiv-Sigve" last="H Varstein">Leiv-Sigve H Varstein</name><affiliation><mods:affiliation>Center of Biotechnology, University of Bergen, Norway</mods:affiliation></affiliation></author><author><name sortKey="Kleppe, Kjell" sort="Kleppe, Kjell" uniqKey="Kleppe K" first="Kjell" last="Kleppe">Kjell Kleppe</name><affiliation><mods:affiliation>Center of Biotechnology, University of Bergen, Norway</mods:affiliation></affiliation></author></analytic><monogr></monogr><series><title level="j">Analytical Biochemistry</title><title level="j" type="abbrev">YABIO</title><idno type="ISSN">0003-2697</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1990">1990</date><biblScope unit="volume">189</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="40">40</biblScope><biblScope unit="page" to="50">50</biblScope></imprint><idno 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scintillation</term><term>Lysate</term><term>Magnetic beads</term><term>Main chain</term><term>Measles</term><term>Measles cells</term><term>Measles virus</term><term>Methoxy phosphoramidites</term><term>Mrna</term><term>Negative control</term><term>Nonspecific binding</term><term>Nucleic</term><term>Nucleic acids</term><term>Nucleoside</term><term>Oligonucleotide</term><term>Oligonucleotide synthesis</term><term>Oligonucleotides</term><term>Plasmid</term><term>Prehybridization</term><term>Radiolabeled oligonucleotide</term><term>Room temperature</term><term>Sandwich hybridization</term><term>Scintillation</term><term>Solid support</term><term>Synthesizer</term><term>Target plasmid</term><term>Uninfected cells</term><term>Urethane group</term></keywords></textClass><langUsage><language ident="en">en</language></langUsage></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Abstract: A novel, rapid, one-step isolation procedure utilizing oligonucleotides end-attached to 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The beads and their captured target nucleic acids were separated after 2 h from the hybridization solution using an external magnet. This procedure was used to isolate measles virus hemagglutinin (H) mRNA from infected cells dissolved in 5 m guanidine thiocyanate. The yield of the specific, undegraded H-mRNA was found to be near the estimated total amount of H-mRNA present in the cells. The magnetic beads were also used for detection of measles virus H-mRNA in a crude cell lysate by sandwich hybridization. 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The beads and their captured target nucleic acids were separated after 2 h from the hybridization solution using an external magnet. This procedure was used to isolate measles virus hemagglutinin (H) mRNA from infected cells dissolved in 5 m guanidine thiocyanate. The yield of the specific, undegraded H-mRNA was found to be near the estimated total amount of H-mRNA present in the cells. The magnetic beads were also used for detection of measles virus H-mRNA in a crude cell lysate by sandwich hybridization. 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Biotechnology, University of Bergen, Norway</affiliation><affiliation>International Centre for Genetic Engineering and Biotechnology, Trieste, Italy</affiliation><affiliation>1Present address: International Centre for Genetic Engineering and Biotechnology, Padriciano 99, I-34012 Trieste, Italy.</affiliation></author><author xml:id="author-0001"><persName><forename type="first">Karl-Henning</forename><surname>Kalland</surname></persName><affiliation>Center of Biotechnology, University of Bergen, Norway</affiliation></author><author xml:id="author-0002"><persName><forename type="first">Bjørn-Ivar</forename><surname>Haukanes</surname></persName><affiliation>Center of Biotechnology, University of Bergen, Norway</affiliation></author><author xml:id="author-0003"><persName><forename type="first">Leiv-Sigve</forename><surname>Håvarstein</surname></persName><affiliation>Center of Biotechnology, University of Bergen, Norway</affiliation></author><author xml:id="author-0004"><persName><forename type="first">Kjell</forename><surname>Kleppe</surname></persName><affiliation>Center of Biotechnology, University of Bergen, Norway</affiliation></author><idno type="istex">F55E88B7AFF080D43CBDABCCCA291DEFEB816599</idno><idno type="ark">ark:/67375/6H6-1BJRCGZZ-0</idno><idno type="DOI">10.1016/0003-2697(90)90041-7</idno><idno type="PII">0003-2697(90)90041-7</idno><idno type="article-id">90900417</idno></analytic><monogr><title level="j">Analytical Biochemistry</title><title level="j" type="abbrev">YABIO</title><idno type="pISSN">0003-2697</idno><idno type="PII">S0003-2697(00)X0602-1</idno><imprint><publisher>ELSEVIER</publisher><date type="published" when="1990"></date><biblScope unit="volume">189</biblScope><biblScope unit="issue">1</biblScope><biblScope unit="page" from="40">40</biblScope><biblScope unit="page" to="50">50</biblScope></imprint></monogr></biblStruct></sourceDesc></fileDesc><profileDesc><creation><date>1990</date></creation><langUsage><language 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The beads and their captured target nucleic acids were separated after 2 h from the hybridization solution using an external magnet. This procedure was used to isolate measles virus hemagglutinin (H) mRNA from infected cells dissolved in 5 m guanidine thiocyanate. The yield of the specific, undegraded H-mRNA was found to be near the estimated total amount of H-mRNA present in the cells. The magnetic beads were also used for detection of measles virus H-mRNA in a crude cell lysate by sandwich hybridization. The experimental detection limit was 630 amol H-mRNA.</p></abstract></profileDesc><revisionDesc><change when="1990">Published</change></revisionDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-1BJRCGZZ-0/fulltext.txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Elsevier, elements deleted: tail"><istex:xmlDeclaration>version="1.0" encoding="UTF-8"</istex:xmlDeclaration><istex:docType PUBLIC="-//ES//DTD journal article DTD version 4.5.2//EN//XML" URI="art452.dtd" name="istex:docType"></istex:docType><istex:document><converted-article version="4.5.2" docsubtype="fla" xml:lang="en"><item-info><jid>YABIO</jid><aid>90900417</aid><ce:pii>0003-2697(90)90041-7</ce:pii><ce:doi>10.1016/0003-2697(90)90041-7</ce:doi><ce:copyright type="unknown" year="1990"></ce:copyright></item-info><head><ce:title>Applications of magnetic beads with covalently attached oligonucleotides in hybridization: Isolation and detection of specific measles virus mRNA from a crude cell lysate</ce:title><ce:author-group><ce:author><ce:given-name>Catrine</ce:given-name><ce:surname>Albretsen</ce:surname><ce:cross-ref refid="AFF1"><ce:sup loc="post">∗</ce:sup></ce:cross-ref><ce:cross-ref refid="AFF2"><ce:sup loc="post">†</ce:sup></ce:cross-ref><ce:cross-ref refid="FN1"><ce:sup loc="post">1</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Karl-Henning</ce:given-name><ce:surname>Kalland</ce:surname><ce:cross-ref refid="AFF1"><ce:sup loc="post">∗</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Bjørn-Ivar</ce:given-name><ce:surname>Haukanes</ce:surname><ce:cross-ref refid="AFF1"><ce:sup loc="post">∗</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Leiv-Sigve</ce:given-name><ce:surname>Håvarstein</ce:surname><ce:cross-ref refid="AFF1"><ce:sup loc="post">∗</ce:sup></ce:cross-ref></ce:author><ce:author><ce:given-name>Kjell</ce:given-name><ce:surname>Kleppe</ce:surname><ce:cross-ref refid="AFF1"><ce:sup loc="post">∗</ce:sup></ce:cross-ref></ce:author><ce:affiliation id="AFF1"><ce:label>a</ce:label><ce:textfn>Center of Biotechnology, University of Bergen, Norway</ce:textfn></ce:affiliation><ce:affiliation id="AFF2"><ce:label>b</ce:label><ce:textfn>International Centre for Genetic Engineering and Biotechnology, Trieste, Italy</ce:textfn></ce:affiliation><ce:footnote id="FN1"><ce:label>1</ce:label><ce:note-para>Present address: International Centre for Genetic Engineering and Biotechnology, Padriciano 99, I-34012 Trieste, Italy.</ce:note-para></ce:footnote></ce:author-group><ce:date-received day="16" month="3" year="1990"></ce:date-received><ce:abstract class="author"><ce:section-title>Abstract</ce:section-title><ce:abstract-sec><ce:simple-para view="all" id="simple-para.0010">A novel, rapid, one-step isolation procedure utilizing oligonucleotides end-attached to magnetic beads (Dynabeads) has been developed. The beads and their captured target nucleic acids were separated after 2 h from the hybridization solution using an external magnet. This procedure was used to isolate measles virus hemagglutinin (H) mRNA from infected cells dissolved in 5 <ce:small-caps>m</ce:small-caps> guanidine thiocyanate. The yield of the specific, undegraded H-mRNA was found to be near the estimated total amount of H-mRNA present in the cells. The magnetic beads were also used for detection of measles virus H-mRNA in a crude cell lysate by sandwich hybridization. The experimental detection limit was 630 amol H-mRNA.</ce:simple-para></ce:abstract-sec></ce:abstract></head></converted-article></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Applications of magnetic beads with covalently attached oligonucleotides in hybridization: Isolation and detection of specific measles virus mRNA from a crude cell lysate</title></titleInfo><titleInfo type="alternative" lang="en" contentType="CDATA"><title>Applications of magnetic beads with covalently attached oligonucleotides in hybridization: Isolation and detection of specific measles virus mRNA from a crude cell lysate</title></titleInfo><name type="personal"><namePart type="given">Catrine</namePart><namePart type="family">Albretsen</namePart><affiliation>Center of Biotechnology, University of Bergen, Norway</affiliation><affiliation>International Centre for Genetic Engineering and Biotechnology, Trieste, Italy</affiliation><affiliation>1Present address: International Centre for Genetic Engineering and Biotechnology, Padriciano 99, I-34012 Trieste, Italy.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Karl-Henning</namePart><namePart type="family">Kalland</namePart><affiliation>Center of Biotechnology, University of Bergen, Norway</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Bjørn-Ivar</namePart><namePart type="family">Haukanes</namePart><affiliation>Center of Biotechnology, University of Bergen, Norway</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Leiv-Sigve</namePart><namePart type="family">Håvarstein</namePart><affiliation>Center of Biotechnology, University of Bergen, Norway</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Kjell</namePart><namePart type="family">Kleppe</namePart><affiliation>Center of Biotechnology, University of Bergen, Norway</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="research-article" displayLabel="Full-length article" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-1JC4F85T-7">research-article</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1990</dateIssued><copyrightDate encoding="w3cdtf">1990</copyrightDate></originInfo><language><languageTerm type="code" authority="iso639-2b">eng</languageTerm><languageTerm type="code" authority="rfc3066">en</languageTerm></language><abstract lang="en">Abstract: A novel, rapid, one-step isolation procedure utilizing oligonucleotides end-attached to magnetic beads (Dynabeads) has been developed. The beads and their captured target nucleic acids were separated after 2 h from the hybridization solution using an external magnet. This procedure was used to isolate measles virus hemagglutinin (H) mRNA from infected cells dissolved in 5 m guanidine thiocyanate. The yield of the specific, undegraded H-mRNA was found to be near the estimated total amount of H-mRNA present in the cells. The magnetic beads were also used for detection of measles virus H-mRNA in a crude cell lysate by sandwich hybridization. The experimental detection limit was 630 amol H-mRNA.</abstract><relatedItem type="host"><titleInfo><title>Analytical Biochemistry</title></titleInfo><titleInfo type="abbreviated"><title>YABIO</title></titleInfo><genre type="journal" authority="ISTEX" authorityURI="https://publication-type.data.istex.fr" valueURI="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</genre><originInfo><publisher>ELSEVIER</publisher><dateIssued encoding="w3cdtf">1990</dateIssued></originInfo><identifier type="ISSN">0003-2697</identifier><identifier type="PII">S0003-2697(00)X0602-1</identifier><part><date>1990</date><detail type="volume"><number>189</number><caption>vol.</caption></detail><detail type="issue"><number>1</number><caption>no.</caption></detail><extent unit="issue-pages"><start>1</start><end>150</end></extent><extent unit="pages"><start>40</start><end>50</end></extent></part></relatedItem><identifier type="istex">F55E88B7AFF080D43CBDABCCCA291DEFEB816599</identifier><identifier type="ark">ark:/67375/6H6-1BJRCGZZ-0</identifier><identifier type="DOI">10.1016/0003-2697(90)90041-7</identifier><identifier type="PII">0003-2697(90)90041-7</identifier><identifier type="ArticleID">90900417</identifier><recordInfo><recordContentSource authority="ISTEX" authorityURI="https://loaded-corpus.data.istex.fr" valueURI="https://loaded-corpus.data.istex.fr/ark:/67375/XBH-HKKZVM7B-M">elsevier</recordContentSource></recordInfo></mods><json:item><extension>json</extension><original>false</original><mimetype>application/json</mimetype><uri>https://api.istex.fr/ark:/67375/6H6-1BJRCGZZ-0/record.json</uri></json:item></metadata><serie></serie></istex></record>Pour manipuler ce document sous Unix (Dilib)
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