Real-time reverse-transcription polymerase chain reaction

Bustin SA, Nolan T. 2004. Pitfalls of quantitative real-time reverse-transcription polymerase chain reaction. J Biomol Tech 15 155-166. 

M.M. Wolf, C.R. Ferguson, J. Ibbotson, S.Fl. Quantitative real-time reverse transcription-polymerase chain reaction analysis of drug metabolizing and cytoprotective... 

G. Smith, R. S. Dawe, C. Clark, A. T. Evans, M. M. Comrie, C. R. Wolf, J. Ferguson, and S. H. Ibbotson, Quantitative real-time reverse transcription-polymerase chain reaction analysis of drug metabolizing and cytoprotective genes in psoriasis and regulation by ultraviolet radiation. J Invest Dermatol 121(2) 390-398 (2003). 

Viale G, Dell Orto P, Biasi MO, et al. Comparative evaluation of an extensive histopathologic examination and a real-time reverse-transcription-polymerase chain reaction assay for mam-maglobin and cytokeratin 19 on axillary sentinel lymph nodes of breast carcinoma patients. Ann Surg. 2008 247 136-142. 

Alcorn, J., Lu, X., Moscow, J. A., and McNamara, P. J. (2002) Transporter gene expression in lactating and nonlactating human mammary epithelial cells using real-time reverse transcription-polymerase chain reaction. J. Pharmacol. Exp. Ther. 303, 487 496. 

Bustin SA. Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J Mol Endocrinol 2000 25 169-193. 

Almeida, A., Thiery, J.P., Magdelenat, H., and Radvanyi, F. (2004) Gene exper-ession analysis by real-time reverse transcription polymerase chain reaction influence of tissue handling. Anal. Biochem. 328 101-108. 

Liu, W. and Saint, D. A. 2002. A new quantitative method of real time reverse transcription polymerase chain reaction assay based on simulation of polymerase chain reaction kinetics. Anal Biochem, 302 52-9. 

Bijwaard, K.E., Fetsch, J.F., Przygodzki, R. et al. (2002) Detection of SYT-SSX fusion transcript in archival synovial sarcoma by Real-time reverse-transcriptase polymerase chain reaction. J Mol Diag, 4, 59-64. 

Reverse transcription-polymerase chain reaction (RT-PCR) has been widely used for the detection of cytokine gene expression in clinical samples (W18, K5). However, conventional RT-PCR only offers a semiquantitative analysis. Recently, the Perkin-Elmer Corporation (Wellesley, MA) developed the TaqMan cytokine gene expression plate for real-time, in vitro quantitative evaluation of a panel of human cytokine gene expression using fluorescence detection. 

Because the 5-HT4 receptors mediate physiological effects in the heart, gut, and CNS (132), splice variants of this receptor are thought to be involved in atrial arrhythmia, irritable bowel syndrome, and neurodegenerative diseases. Medhurst et al. (132) used TaqMan real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR) to investigate the mRNA distribution of 5-HT4 receptor C-terminal splice variants in multiple human CNS and peripheral tissues. They... 

S. J. Wall and D. R. Edwards, Quantitative reverse transcription-polymerase chain reaction (RT-PCR) a comparison of primer-dropping, competitive, and real-time RT-PCRs. Anal Biochem 300(2) 269-273 (2002). 

Ueno, H., Yoshida, K., Hirai, T., Kono, F., Kambe, M., and Toge, T., Quantitative detection of carcinoembryonic antigen messenger RNA in the peritoneal cavity of gastric cancer patients by real-time quantitative reverse transcription polymerase chain reaction. Anticancer Res. 23, 1701-1708 (2003). 

Related mRNAs encoding various proteins can be detected by different types of in situ hybridization. For this method, the number of specific mRNAs detectable per tumor sample is limited. However, the advantage of in situ hybridization is the same as in immunohistochemistry where the morphology of the tumor is still visible. Specific mRNA-species can be detected by northern blot, nuclease protection assay or reverse transcription (RT) combined with polymerase chain reaction (PCR). Using the modern real-time PCR protocols, reliable quantification of PCR targets is possible. A more complex approach is possible by using the micro-array technology, where hundreds or even more of mRNAs can be detected simultaneously in a semi-quantitative fashion. 

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