[PDF][PDF] Short technical report processing of gene expression data generated by quantitative real-time RT-PCR

PY Muller, H Janovjak, AR Miserez… - …, 2002 - gene-quantification.com
PY Muller, H Janovjak, AR Miserez, Z Dobbie
Biotechniques, 2002gene-quantification.com
Real-time quantitative PCR represents a highly sensitive and powerful technique for the
quantitation of nucleic acids. It has a tremendous potential for the high-throughput analysis
of gene expression in research and routine diagnostics. However, the major hurdle is not the
practical performance of the experiments themselves but rather the efficient evaluation and
the mathematical and statistical analysis of the enormous amount of data gained by this
technology, as these functions are not included in the software provided by the …
Abstract
Real-time quantitative PCR represents a highly sensitive and powerful technique for the quantitation of nucleic acids. It has a tremendous potential for the high-throughput analysis of gene expression in research and routine diagnostics. However, the major hurdle is not the practical performance of the experiments themselves but rather the efficient evaluation and the mathematical and statistical analysis of the enormous amount of data gained by this technology, as these functions are not included in the software provided by the manufacturers of the detection systems. In this work, we focus on the mathematical evaluation and analysis of the data generated by real-time quantitative PCR, the calculation of the final results, the propagation of experimental variation of the measured values to the final results, and the statistical analysis. We developed a Microsoft® Excel®-based software application coded in Visual Basic for Applications, called Q-Gene, which addresses these points. Q-Gene manages and expedites the planning, performance, and evaluation of real-time quantitative PCR experiments, as well as the mathematical and statistical analysis, storage, and graphical presentation of the data. The Q-Gene software application is a tool to cope with complex realtime quantitative PCR experiments at a high-throughput scale and considerably expedites and rationalizes the experimental setup, data analysis, and data management while ensuring highest reproducibility.
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