A novel medium throughput quantitative competitive PCR technology to simultaneously measure mRNA levels from multiple genes
A novel medium throughput quantitative competitive PCR technology to simultaneously measure mRNA levels from multiple genes
There is a great demand for technologies to simultaneously measure mRNA levels from multiple genes. Here we report a new quantitative competitive PCR technology and demonstrate simultaneous quantification of mRNA from multiple genes. First, a sequential 2-fold dilution series containing equal amounts of gene-specific standard DNAs for 10-12 genes is prepared. Second, the serially diluted standard DNAs are individually added to equal amounts of tissue-derived cDNA and amplified with gene-specific primers for 19-26 PCR cycles. Each gene/standard DNA pair is amplified individually. All amplified DNA products (n = 80) are resolved by one microplate array diagonal gel electrophoresis using 5% polyacrylamide. Changes in mRNA levels of approximately 15% can be detected by this technology. The mRNA levels from 10-12 genes were simultaneously quantified. mRNA levels were compared in RNA samples from rat liver, kidney and skeletal muscle. This quick, specific, sensitive, reproducible and yet inexpensive technique is ideal for simultaneously studying co-ordinate changes in mRNA levels from multiple genes.
research support, dna, non-u.s.gov't, genetic, disease, rats, analysis, kidney, liver, reference standards, skeletal, report, polyacrylamide gel, messenger, polymerase chain reaction, animals
e20-[10pp]
Zhang, Junlong
68a8fa77-c5db-4b34-aa9a-fbad6860155f
Day, Ian N.M.
b749b30a-1f4c-40eb-af0e-a50427388b39
Byrne, Christopher D.
1370b997-cead-4229-83a7-53301ed2a43c
14 January 2002
Zhang, Junlong
68a8fa77-c5db-4b34-aa9a-fbad6860155f
Day, Ian N.M.
b749b30a-1f4c-40eb-af0e-a50427388b39
Byrne, Christopher D.
1370b997-cead-4229-83a7-53301ed2a43c
Zhang, Junlong, Day, Ian N.M. and Byrne, Christopher D.
(2002)
A novel medium throughput quantitative competitive PCR technology to simultaneously measure mRNA levels from multiple genes.
Nucleic Acids Research, 30 (5), .
(doi:10.1093/nar/30.5.e20).
Abstract
There is a great demand for technologies to simultaneously measure mRNA levels from multiple genes. Here we report a new quantitative competitive PCR technology and demonstrate simultaneous quantification of mRNA from multiple genes. First, a sequential 2-fold dilution series containing equal amounts of gene-specific standard DNAs for 10-12 genes is prepared. Second, the serially diluted standard DNAs are individually added to equal amounts of tissue-derived cDNA and amplified with gene-specific primers for 19-26 PCR cycles. Each gene/standard DNA pair is amplified individually. All amplified DNA products (n = 80) are resolved by one microplate array diagonal gel electrophoresis using 5% polyacrylamide. Changes in mRNA levels of approximately 15% can be detected by this technology. The mRNA levels from 10-12 genes were simultaneously quantified. mRNA levels were compared in RNA samples from rat liver, kidney and skeletal muscle. This quick, specific, sensitive, reproducible and yet inexpensive technique is ideal for simultaneously studying co-ordinate changes in mRNA levels from multiple genes.
This record has no associated files available for download.
More information
Published date: 14 January 2002
Keywords:
research support, dna, non-u.s.gov't, genetic, disease, rats, analysis, kidney, liver, reference standards, skeletal, report, polyacrylamide gel, messenger, polymerase chain reaction, animals
Identifiers
Local EPrints ID: 60460
URI: http://eprints.soton.ac.uk/id/eprint/60460
ISSN: 0305-1048
PURE UUID: 656fb0df-0050-4af1-b300-7bdb63ce42e6
Catalogue record
Date deposited: 08 Sep 2008
Last modified: 16 Mar 2024 03:07
Export record
Altmetrics
Contributors
Author:
Junlong Zhang
Author:
Ian N.M. Day
Download statistics
Downloads from ePrints over the past year. Other digital versions may also be available to download e.g. from the publisher's website.
View more statistics
