Quantitative detection of messenger RNA by solution hybridization and enzyme immunoassay

Francois Coutlee; Elizabeth A. Rubalcaba; Raphael P. Viscidi; James E. Gern; Patrick A. Murphy; Howard M. Lederman

Quantitative detection of messenger RNA by solution hybridization and enzyme immunoassay

Francois Coutlee, Elizabeth A. Rubalcaba, Raphael P. Viscidi, James E. Gern, Patrick A. Murphy, Howard M. Lederman

School of Medicine

Research output: Contribution to journal › Article › peer-review

14 Scopus citations

Abstract

A novel nucleic acid detection technique is described for the quantitative measurement of eukaryotic mRNA in biological samples. The procedure involves two steps: a hybridization reaction in solution with a biotinylated cDNA probe, and a conventional enzyme immunoassay that uses a monoclonal antibody for DNA·RNA hybrids to detect the specific mRNA·cDNA complexes. The method has comparable sensitivity to ³²P-based methods and yields results that are quantitative and highly reproducible. Furthermore, the test can be performed using unfractionated cytoplasm without the need for extraction with organic solvents. This technique provides a rapid and quantitative method for studying changes in cellular mRNA levels, and it is suitable for testing large numbers of samples.

Original language	English (US)
Pages (from-to)	11601-11604
Number of pages	4
Journal	Journal of Biological Chemistry
Volume	265
Issue number	20
State	Published - Jul 15 1990

ASJC Scopus subject areas

Biochemistry
Molecular Biology
Cell Biology

Cite this

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title = "Quantitative detection of messenger RNA by solution hybridization and enzyme immunoassay",

abstract = "A novel nucleic acid detection technique is described for the quantitative measurement of eukaryotic mRNA in biological samples. The procedure involves two steps: a hybridization reaction in solution with a biotinylated cDNA probe, and a conventional enzyme immunoassay that uses a monoclonal antibody for DNA·RNA hybrids to detect the specific mRNA·cDNA complexes. The method has comparable sensitivity to 32P-based methods and yields results that are quantitative and highly reproducible. Furthermore, the test can be performed using unfractionated cytoplasm without the need for extraction with organic solvents. This technique provides a rapid and quantitative method for studying changes in cellular mRNA levels, and it is suitable for testing large numbers of samples.",

author = "Francois Coutlee and Rubalcaba, {Elizabeth A.} and Viscidi, {Raphael P.} and Gern, {James E.} and Murphy, {Patrick A.} and Lederman, {Howard M.}",

year = "1990",

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T1 - Quantitative detection of messenger RNA by solution hybridization and enzyme immunoassay

AU - Coutlee, Francois

AU - Rubalcaba, Elizabeth A.

AU - Viscidi, Raphael P.

AU - Gern, James E.

AU - Murphy, Patrick A.

AU - Lederman, Howard M.

PY - 1990/7/15

Y1 - 1990/7/15

N2 - A novel nucleic acid detection technique is described for the quantitative measurement of eukaryotic mRNA in biological samples. The procedure involves two steps: a hybridization reaction in solution with a biotinylated cDNA probe, and a conventional enzyme immunoassay that uses a monoclonal antibody for DNA·RNA hybrids to detect the specific mRNA·cDNA complexes. The method has comparable sensitivity to 32P-based methods and yields results that are quantitative and highly reproducible. Furthermore, the test can be performed using unfractionated cytoplasm without the need for extraction with organic solvents. This technique provides a rapid and quantitative method for studying changes in cellular mRNA levels, and it is suitable for testing large numbers of samples.

AB - A novel nucleic acid detection technique is described for the quantitative measurement of eukaryotic mRNA in biological samples. The procedure involves two steps: a hybridization reaction in solution with a biotinylated cDNA probe, and a conventional enzyme immunoassay that uses a monoclonal antibody for DNA·RNA hybrids to detect the specific mRNA·cDNA complexes. The method has comparable sensitivity to 32P-based methods and yields results that are quantitative and highly reproducible. Furthermore, the test can be performed using unfractionated cytoplasm without the need for extraction with organic solvents. This technique provides a rapid and quantitative method for studying changes in cellular mRNA levels, and it is suitable for testing large numbers of samples.

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M3 - Article

C2 - 2195023

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JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

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ER -

Quantitative detection of messenger RNA by solution hybridization and enzyme immunoassay

Abstract

ASJC Scopus subject areas

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