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STAYING CURRENT: TECHNOLOGY

The power of real-time PCR

¹ Department of Physiology Touchstone Center for Diabetes Research, University of Texas Southwestern Medical Center, Dallas, Texas
² Department of Internal Medicine, Touchstone Center for Diabetes Research, University of Texas Southwestern Medical Center, Dallas, Texas

Address for reprint requests and other correspondence: J. J. Repa, Univ. of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Y8.212B, Dallas, TX 75390-8854 (E-mail: joyce.repa{at}utsouthwestern.edu)

Abstract

In recent years, real-time polymerase chain reaction (PCR) has emerged as a robust and widely used methodology for biological investigation because it can detect and quantify very small amounts of specific nucleic acid sequences. As a research tool, a major application of this technology is the rapid and accurate assessment of changes in gene expression as a result of physiology, pathophysiology, or development. This method can be applied to model systems to measure responses to experimental stimuli and to gain insight into potential changes in protein level and function. Thus physiology can be correlated with molecular events to gain a better understanding of biological processes. For clinical molecular diagnostics, real-time PCR can be used to measure viral or bacterial loads or evaluate cancer status. Here, we discuss the basic concepts, chemistries, and instrumentation of real-time PCR and include present applications and future perspectives for this technology in biomedical sciences and in life science education.

Key words: quantitative real-time polymerase chain reaction; method; review; adiponectin receptor

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