The reverse transcription polymerase chain reaction (RT-PCR) is the most sensitive method for the detection of low-abundance mRNA, often obtained from limited tissue samples. However, it is a complex technique, there are substantial problems associated with its true sensitivity, reproducibility and specificity and, as a quantitative method, it suffers from the problems inherent in PCR. The recent introduction of fluorescence-based kinetic RT-PCR procedures significantly simplifies the process of producing reproducible quantification of mRNAs and promises to overcome these limitations. Nevertheless, their successful application depends on a clear understanding of the practical problems, and careful experimental design, application and validation remain essential for accurate quantitative measurements of transcription. This review discusses the technical aspects involved, contrasts conventional and kinetic RT-PCR methods for quantitating gene expression and compares the different kinetic RT-PCR systems. It illustrates the usefulness of these assays by demonstrating the significantly different levels of transcription between individuals of the housekeeping gene family, glyceraldehyde-3-phosphate-dehydrogenase (GAPDH).

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				K. Kamio, X. Liu, H. Sugiura, S. Togo, T. Kobayashi, S. Kawasaki, X. Wang, L. Mao, Y. Ahn, C. Hogaboam, et al. Prostacyclin Analogs Inhibit Fibroblast Contraction of Collagen Gels through the cAMP-PKA Pathway Am. J. Respir. Cell Mol. Biol., July 1, 2007; 37(1): 113 - 120. [Abstract] [Full Text] [PDF]

				X. Liu, A. M. Das, J. Seideman, D. Griswold, C. N. Afuh, T. Kobayashi, S. Abe, Q. Fang, M. Hashimoto, H. Kim, et al. The CC Chemokine Ligand 2 (CCL2) Mediates Fibroblast Survival through IL-6 Am. J. Respir. Cell Mol. Biol., July 1, 2007; 37(1): 121 - 128. [Abstract] [Full Text] [PDF]

				A. Amaral, J. Ramalho-Santos, and J. C. St John The expression of polymerase gamma and mitochondrial transcription factor A and the regulation of mitochondrial DNA content in mature human sperm Hum. Reprod., June 1, 2007; 22(6): 1585 - 1596. [Abstract] [Full Text] [PDF]

				M. Cronin, C. Sangli, M.-L. Liu, M. Pho, D. Dutta, A. Nguyen, J. Jeong, J. Wu, K. C. Langone, and D. Watson Analytical Validation of the Oncotype DX Genomic Diagnostic Test for Recurrence Prognosis and Therapeutic Response Prediction in Node-Negative, Estrogen Receptor-Positive Breast Cancer Clin. Chem., June 1, 2007; 53(6): 1084 - 1091. [Abstract] [Full Text] [PDF]

				X. Jiang, K. M. Saw, A. Eaves, and C. Eaves Instability of BCR-ABL Gene in Primary and Cultured Chronic Myeloid Leukemia Stem Cells J Natl Cancer Inst, May 2, 2007; 99(9): 680 - 693. [Abstract] [Full Text] [PDF]

				M. Shemesh, A. Tam, and D. Steinberg Differential gene expression profiling of Streptococcus mutans cultured under biofilm and planktonic conditions Microbiology, May 1, 2007; 153(5): 1307 - 1317. [Abstract] [Full Text] [PDF]

				N. A. Janovick-Guretzky, H. M. Dann, D. B. Carlson, M. R. Murphy, J. J. Loor, and J. K. Drackley Housekeeping Gene Expression in Bovine Liver is Affected by Physiological State, Feed Intake, and Dietary Treatment J Dairy Sci, May 1, 2007; 90(5): 2246 - 2252. [Abstract] [Full Text] [PDF]

				H. Francois, C. Facemire, A. Kumar, L. Audoly, B. Koller, and T. Coffman Role of Microsomal Prostaglandin E Synthase 1 in the Kidney J. Am. Soc. Nephrol., May 1, 2007; 18(5): 1466 - 1475. [Abstract] [Full Text] [PDF]

				C. E. Wood, T. C. Register, and J.M. Cline Soy isoflavonoid effects on endogenous estrogen metabolism in postmenopausal female monkeys Carcinogenesis, April 1, 2007; 28(4): 801 - 808. [Abstract] [Full Text] [PDF]

				J. Chen, F. F. Kadlubar, and J. Z. Chen DNA supercoiling suppresses real-time PCR: a new approach to the quantification of mitochondrial DNA damage and repair Nucleic Acids Res., March 13, 2007; (2007) gkm010v2. [Abstract] [Full Text] [PDF]

				K. T. Beggs, K. A. Glendining, N. M. Marechal, V. Vergoz, I. Nakamura, K. N. Slessor, and A. R. Mercer Queen pheromone modulates brain dopamine function in worker honey bees PNAS, February 13, 2007; 104(7): 2460 - 2464. [Abstract] [Full Text] [PDF]