期刊
MOLECULAR AND CELLULAR PROBES
卷 18, 期 1, 页码 45-50出版社
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD
DOI: 10.1016/j.mcp.2003.09.001
关键词
quantitative polymerase chain reaction; real-time reverse transcription-polymerase chain reaction; gene expression; housekeeping genes; ubiquitin; beta-actin; GAPDH; 18S rRNA
Real-time reverse transcription-polymerase chain reaction (RT-PCR) is currently considered the most sensitive method to study low abundance gene expression. Since comparison of gene expression levels in various tissues is often the purpose of an experiment, we studied a tissue-linked effect on nucleic acid amplification. Based on the raw data generated by a LightCycler instrument, we propose a descriptive mathematical model of PCR amplification. This model allowed us to study amplification kinetics of four common housekeeping genes in total RNA samples derived from various bovine tissues. We observed that unknown tissue-specific factors can influence amplification kinetics but this affect can be ameliorated, in part, by appropriate primer selection. (C) 2003 Elsevier Ltd. All rights reserved.
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