Expression measurement of many genes simultaneously by quantitative RT-PCR using standardized mixtures of competitive templates

American Journal of Respiratory Cell and Molecular Biology
James C WilleyJeffrey P DeMuth

Abstract

Progress toward complete sequencing of all human genes through the Human Genome Project has already resulted in a need for methods that allow quantitative expression measurement of multiple genes simultaneously. It is increasingly recognized that relative measurement of multiple genes will provide more mechanistic information regarding cell pathophysiology than measurement of individual genes one by one or by methods that do not allow direct intergene comparison. In this study, previously described quantitative reverse transcription-polymerase chain reaction methods were modified in an effort to provide a rapid, simple method for this purpose. Internal standard competitive templates (CTs) were prepared for each gene and were combined in a single solution containing CTs for more than 40 genes at defined concentrations relative to one another. Any subsequent dilution of the CT mixture did not alter the relationship of one CT to another. Because the same CT standard solution or a dilution of it was used in all experiments, data obtained from different experiments were easily compared. The use of multiple CT mixtures with different housekeeping gene to target gene ratios provided a linear dynamic range spanning the range of express...Continue Reading

References

Apr 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·G GillilandH F Bunn
Oct 20, 1995·Science·V E VelculescuK W Kinzler
Sep 1, 1993·Analytical Biochemistry·M J ApostolakosJ C Willey
Feb 25, 1993·Nucleic Acids Research·F S CeliA R Shuldiner
Mar 1, 1996·American Journal of Respiratory Cell and Molecular Biology·J C WilleyK Cairns
Dec 1, 1996·Nature Biotechnology·D J LockhartE L Brown
Jul 3, 1998·American Journal of Respiratory Cell and Molecular Biology·J P DeMuthJ C Willey
Jul 3, 1998·American Journal of Respiratory Cell and Molecular Biology·J P DeMuthJ C Willey

❮ Previous
Next ❯

Citations

Jul 13, 2002·International Journal of Cancer. Journal International Du Cancer·Kiyoshi ItoHironobu Sasano
Dec 5, 2012·Analytical and Bioanalytical Chemistry·Jia LiLi Li
Mar 15, 2003·Clinica Chimica Acta; International Journal of Clinical Chemistry·John R PetersenDeborah A Payne
Mar 12, 2003·European Journal of Cancer : Official Journal for European Organization for Research and Treatment of Cancer (EORTC) [and] European Association for Cancer Research (EACR)·C L van der WiltG J Peters
Oct 11, 2003·Transplant Immunology·Paul J GibbsW Martin Howell
Feb 8, 2003·American Journal of Respiratory and Critical Care Medicine·Satoshi SuzukiZygmund S Krozowski
Jul 3, 1998·American Journal of Respiratory Cell and Molecular Biology·J P DeMuthJ C Willey
Jul 3, 1998·American Journal of Respiratory Cell and Molecular Biology·J P DeMuthJ C Willey
Jul 3, 1998·American Journal of Respiratory Cell and Molecular Biology·W N Rom
Oct 29, 2005·BMC Genomics·Vigdis NygaardEivind Hovig
Jul 22, 2009·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·Anirban P MitraRichard J Cote
Jul 13, 2002·International Journal of Cancer. Journal International Du Cancer·Martin VondracekRoland C Grafström
Oct 30, 2010·International Journal of Hygiene and Environmental Health·Sirilak WongvijitsukMathuros Ruchirawat
Jul 29, 2006·International Journal of Hygiene and Environmental Health·Wanwisa SudprasertMathuros Ruchirawat
Oct 19, 2005·Journal of Virological Methods·Søren GroveBirgit Helene Dannevig
Jun 11, 2002·Biochemical and Biophysical Research Communications·Erin L CrawfordJames C Willey
Feb 22, 2001·Experimental Eye Research·T UdonoS Shibahara
Mar 27, 2001·Japanese Journal of Cancer Research : Gann·N ArigaH Sasano
Sep 27, 2001·Japanese Journal of Cancer Research : Gann·J I AkahiraH Sasano
Jul 28, 2013·Gene Regulation and Systems Biology·Thomas M BlomquistJames C Willey
Dec 2, 2004·Nucleosides, Nucleotides & Nucleic Acids·M de BruinG J Peters
Aug 26, 2006·Applied Immunohistochemistry & Molecular Morphology : AIMM·Clive R Taylor
Jan 8, 2003·Journal of Environmental Science and Health. Part C, Environmental Carcinogenesis & Ecotoxicology Reviews·Farid E Ahmed
Jul 16, 2008·Molecular and Cellular Biochemistry·Elaine M HurtMichel M Sanders

❮ Previous
Next ❯

Related Concepts

Related Feeds

American Thoracic Association Journals

Discover the latest respiratory research published by the journals from the American Thoracic Society.