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 : 004A76 ( Main/Exploration ); précédent : 004A75; suivant : 004A77Applications 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 [Norvège, Italie] ; Karl-Henning Kalland [Norvège] ; Bj Rn-Ivar Haukanes [Norvège] ; Leiv-Sigve H Varstein [Norvège] ; Kjell Kleppe [Norvège]Source :
- 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
Affiliations:
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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 type="ISSN">0003-2697</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0003-2697</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="Teeft" xml:lang="en"><term>Albretsen</term><term>Ammonia</term><term>Ammonolysis</term><term>Aqueous ammonia</term><term>Autoradiography</term><term>Bead</term><term>Bead surface</term><term>Beads</term><term>Blot analysis</term><term>Blot filter</term><term>Cdna</term><term>Cell lysate</term><term>Cellulose nitrate filter hybridization</term><term>Cellulose nitrate filters</term><term>Crude cell lysate</term><term>Detection limit</term><term>Different amounts</term><term>Different linker lengths</term><term>Eluate</term><term>External magnet</term><term>First wash</term><term>Guanidine thiocyanate</term><term>Guscn</term><term>Herring sperm</term><term>Hmrna</term><term>Hybridization</term><term>Hybridization solution</term><term>Linker</term><term>Linker length</term><term>Liquid 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 magnetic beads (Dynabeads) has been developed. 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