Inhibition of real-time RT-PCR quantification due to tissue-specific contaminants.

• 全文链接
• 请求原文

作   者：

Tichopad A;  Didier A;  Pfaffl MW; 

作者机构：

Institute of Physiology;  Germany.;  FML-Weihenstephan;  Technical University of Munich;  Center of Life and Food Science; 

关键词：

逆转录聚合酶链反应;  kinetic characteristics;  Tissues;  Reverse Transcriptase Polymerase Chain Reaction;  组织; 

期刊名称：

Molecular and Cellular Probes: The Location, Diagnosis and Monitoring of Disease by Specific Molecules and Cell Lines

i s s n：

0890-8508

年卷期：

2004 年 18 卷 1 期

页   码：

45-50

页   码：

摘   要：

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.