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  Quantitative real-time PCR-based analysis of gene expression

Jozefczuk, J., & Adjaye, J. (2011). Quantitative real-time PCR-based analysis of gene expression. Methods in Enzymology, 500, 99-109. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/21943894.

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Jozefczuk, J.1, Author              
Adjaye, J.1, Author              
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1Molecular Embryology and Aging (James Adjaye), Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1479654              

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Free keywords: Algorithms; DNA Primers/chemistry; Embryonic Stem Cells/metabolism; Gene Expression Profiling/*methods; Humans; RNA, Messenger/isolation & purification/metabolism; Real-Time Polymerase Chain Reaction/*methods; Reverse Transcription
 Abstract: Quantitative real-time polymerase chain reaction (QRT-PCR) has become an extensively applied technique. It enables quantitative analyses of gene expression applicable to basic molecular biology, medicine, and diagnostics. Nowadays, it is broadly used to describe messenger RNA (mRNA) expression patterns and to compare the relative levels of mRNA within distinct biological samples. The scope of the QRT-PCR technique makes it applicable across a wide range of experimental conditions and allows experimental comparison between normal and abnormal tissue. Most importantly, this technique enables additional independent confirmation of microarray or next generation sequencing (NGS)-based results. An inherent advantage of QRT-PCR is the large dynamic range, remarkable sensitivity, and sequence-specificity. We provide a detailed step by step guide to the principles underlying a successful QRT-PCR experiment.

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 Dates: 2011
 Publication Status: Published in print
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 Rev. Type: -
 Identifiers: eDoc: 584705
URI: http://www.ncbi.nlm.nih.gov/pubmed/21943894
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Title: Methods in Enzymology
Source Genre: Journal
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Pages: - Volume / Issue: 500 Sequence Number: - Start / End Page: 99 - 109 Identifier: ISSN: 1557-7988 (Electronic) 0076-6879 (Linking)