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Wolf, Hans J. ; Leser, Ulrike ; Haus, M. ; Gu, S. Y. ; Pathmanathan, R.

Sandwich nucleic acid hybridization: a method with a universally usable labeled probe for various specific tests

Wolf, Hans J., Leser, Ulrike, Haus, M., Gu, S. Y. and Pathmanathan, R. (1986) Sandwich nucleic acid hybridization: a method with a universally usable labeled probe for various specific tests. Journal of virological methods 13 (1), pp. 1-8.

Date of publication of this fulltext: 20 Apr 2011 12:43
Article
DOI to cite this document: 10.5283/epub.20555


Abstract

Nucleic acid hybridization is widely used for scientific applications but essentially restricted to specialized laboratories. The use of recombinant m 13 phages as hybridization probes (Hu and Messing (1980) Gene 17, 271; Messing (1983) Methods Enzymol. 101, 20) offers a considerable advantage over the commonly used recombinant plasmids as the preparation of the DNA probe is very simple and it ...

Nucleic acid hybridization is widely used for scientific applications but essentially restricted to specialized laboratories. The use of recombinant m 13 phages as hybridization probes (Hu and Messing (1980) Gene 17, 271; Messing (1983) Methods Enzymol. 101, 20) offers a considerable advantage over the commonly used recombinant plasmids as the preparation of the DNA probe is very simple and it can easily be labeled directly, e.g. with isotopes with long half-life like 125I (Commerford (1971) Biochemistry 10, 11 (1983); Gu et al. (1983) Cancer (China) 2, 129; Han and Harding (1983) Nucleic Acids Res. 11, 14) and used for hybridization. However, as the application of nucleic acid hybridization for diagnostic and epidemiological purposes becomes almost unavoidable, the logistic problems of keeping numerous individually labeled hybridization probes increase considerably and may reach prohibitory levels in less well-equipped laboratories. In a new sandwich technique, the first step involves hybridization with an unlabeled recombinant m 13 DNA carrying an insert of the desired specificity. In a second step a universally usable labeled probe directed against the m 13 part of the recombinant phage DNA is applied. This reduces considerably the problems of preparing and keeping multiple labeled probes in stock.



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Details

Item typeArticle
Journal or Publication TitleJournal of virological methods
Publisher:Elsevier
Volume:13
Number of Issue or Book Chapter:1
Page Range:pp. 1-8
Date1986
InstitutionsMedicine > Lehrstuhl für Medizinische Mikrobiologie und Hygiene
Identification Number
ValueType
3722306PubMed ID
10.1016/0166-0934(86)90066-2DOI
Classification
NotationType
Bacteriophages/geneticsMESH
Base SequenceMESH
DNA, Viral/analysisMESH
Iodine RadioisotopesMESH
Nucleic Acid HybridizationMESH
Dewey Decimal Classification600 Technology > 610 Medical sciences Medicine
StatusPublished
RefereedUnknown
Created at the University of RegensburgUnknown
URN of the UB Regensburgurn:nbn:de:bvb:355-epub-205552
Item ID20555

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