Reverse-transcription polymerase chain reaction

Mies C. A simple, rapid method for isolating RNA from paraffin-embedded tissues for reverse transcription-polymerase chain reaction (RT-PCR). /. Histochem. Cytochem. 1994 42 811-813. 

Godfrey TE, Kim S-H, Chavira M, et al. Quantitative mRNA expression analysis from formalin-fixed, paraffin-embedded tissues using 5 nuclease quantitative reverse transcription-polymerase chain reaction. J. Mol. Diagn. 2000 2 84-91. 

Macabeo-Ong M, Ginzinger DG, Dekker N, et al. Effect of duration of fixation on quantitative reverse transcription polymerase chain reaction analyses. Mod. Pathol. 2002 15 979-987. 

Hamoud MM, Villegas P, Williams SM. Detection of infectious bursal disease virus from formalin-fixed paraffin-embedded tissue by immunohistochemistry and realtime reverse transcription-polymerase chain reaction. J. Vet. Diagn. Invest. 2007 19 35—42. 

Rapid antigen and point-of-care tests, direct fluorescence antibody test, and the reverse-transcription polymerase chain reaction assay may be used for rapid detection of virus. 

Cytokine profiling has also been measured as a function of changes in cytokine mRNA expression using either reverse transcription polymerase chain reaction (RT-PCR) [87, 91-93] or ribonuclease protection assay (RPA) [94-97], Measurement of cytokine transcripts by RT-PCR revealed that prolonged exposure to TMA induced increased levels of IL-4 mRNA expression compared with treatment with DNCB [87,92-93]. However, expression of the type 1 cytokine IFN-y by DNCB-activated LNC was variable and failed to discriminate between contact and respiratory allergens [87,91,93). A similar profile was observed for freshly isolated tissue analyzed by RPA. This somewhat less... 

Hayashi, M. et al., Assessment of preferential Thl or Th2 induction by low-molecular-weight compounds using a reverse transcription-polymerase chain reaction method Comparison of two mouse strains, C57BL/6 and BALB/c. Toxicol. Appl. Pharmacol., 177, 38,2001. 

Y1. Young, K. K. Y., Resnick, R. M., and Myers, T. W., Detection of hepatitis C virus RNA by a combined reverse transcription-polymerase chain reaction assay. J. Clin. Microbiol. 31, 882-886 (1993). 

List of Abbreviations PCR, polymerase chain reaction RT-PCR, reverse transcription polymerase chain reaction DNA, deoxyribonucleic acid RNA, ribonucleic acid RNase, ribonuclease mRNA, messenger RNA GABAa, y-aminobutyric acid type A cRNA, copy RNA dNTPs, deoxy nucleoside triphosphates MMLV, Mouse Moloney murine leukemia vims RT, reverse transcriptase bp, base pair Tm, melting temperature DEPC, diethylpyrocarbonate OD, optical density mL, milliliter SA-PMPs, streptavidin paramagnetic particles dT, deoxy thymidine DTT, dithiothreitol DNase, deoxyribonuclease RNasin, ribonuclease inhibitor UV, ultraviolet TBE, Tris-borate, 1 mM EDTA EDTA, ethylenediaminetetraacetic acid Buffer RET, guanidium thiocyanate lysis buffer PBS, phosphate buffered saline NT2, Ntera 2 neural progenitor cells... 

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

Scherm, B., Palomba, M., Serra, D., Marcello, A., and Megheli, Q. (2005). Detection of transcripts of the aflatoxin genes aflD, aflO, and aflP by reverse transcription-polymerase chain reaction allows differentiation of alfatoxin-producing and non-producing isolates of Aspergillus flavus and Aspergillus parasiticus. Int. J. Food Microbiol. 98, 201-210. 

PTT is based on a combination of reverse-transcription polymerase chain reaction (RT-PCR) and linked in vitro transcription and translation (Fig. 3C). This combination of procedures can selectively detect translation-terminating or nonsense mutations. Unfortunately, it does not find missense mutations, which may be etiologic, depending upon location. 

Yajima, T. Yagihashi, A. Kameshima, H. Furuya, D. Kobayashi, D. Hirata, K. Watanabe, N. Establishment of quantitative reverse transcription-polymerase chain reaction assays for human telomerase-associated genes. Clin. Chim. Acta 2000, 290(2), 117-127. 

Reverse transcription-polymerase chain reaction. Thin layer chromatography. 

The first binding studies on solubilized membranes from rat brain demonstrated the presence of a low affinity adenosine receptor with characteristics of the A3 subtype (Oliveira et al. 1991). However, in situ hybridization studies in the rat indicated the presence of A3AR mRNA only in the testis (Meyerhof et al. 1991a Zhou et al. 1992 Rivkees 1994) and not in the CNS (Rivkees et al. 2000). Similarly, no expression of A3AR in the brain of mice or in the hippocampi of humans was detected (Rivkees et al. 2000). However, by reverse transcription-polymerase chain reaction (RT-PCR)... 

Conventionally, the variants are characterized by coamplification with wild-type sequences using reverse transcription polymerase chain reaction (RT-PCR). However, this approach focuses on small regions of the known wild-type mRNA. Because of this threshold detection, spliced transcripts expressed at low levels may fall below the threshold of detection. To avoid this and other limitations of the conventional RT-PCR technique, the targeted amplification method can be used (Poola et al., 2000). This method involves the targeted amplification of the alternatively spliced molecules as separate gene populations using specific primers designed for the alternative splice junctions, without coamplification of wild-type molecules. 

A reliable method of measuring the ER content in human breast cancer is important for optimal treatment and a qualified estimate of the recurrence-free survival of the patient. The majority of the studies on the expression of ERs, especially ER 3 in human tissues, have been accomplished using RNA techniques such as reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization. Although the RT-PCR method is an effective tool to describe the presence of a particular gene in the tissue, this approach does not indicate the specific cell that expresses the gene. 

Helguera, P. R., Taborda, R., Docampo, D. M., and Ducasse, D. A. (2001) Immunocapture reverse transcription-polymerase chain reaction combined with nested PCR greatly increases the detection of Prunus necrotic ring spot virus in the peach. J. Virol. Methods 95, 93-100. 

Zhou et al. [175] described the determination of severe acute respiratory syndrome (SARS) coronavirus by a microfluidic chip system. The unit included an LIF microfluidic chip analyzer, a glass microchip for both PCR and capillary electrophoresis, a chip thermal cycler based on dual Peltier thermoelectric elements, a reverse transcription-polymerase chain reaction (RT-PCR) SARS diagnostic kit, and a DNA electrophoretic sizing kit. According to the authors, the system allowed efficient DNA amplification of the SARS coronavirus followed by electrophoretic sizing and detection on the same chip. 

Single cell reverse transcription - polymerase chain reaction detected D2 and D5 but not D3/4 and little Di receptor mRNA in rat striatal cholinergic intemeurons. The D2 receptors coupled to Gi/o and reduced somatic N-type Ca2+ currents (Yan et al. 1997), thus providing a cellular mechanism for the reduction of acetylcholine release by D2 receptors located on cholinergic terminals. 

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. 

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