SAS programs for real-time RT-PCR having multiple independent samples.

Peyton Cook; Chunxiao Fu; Morgen Hickey; Eun-Soo Han; Kenton S Miller

doi:10.2144/04376bin02

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SAS programs for real-time RT-PCR having multiple independent samples.

Peyton Cook·Chunxiao Fu·Morgen Hickey·Eun-Soo Han·Kenton S Miller

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Abstract

Relative real-time reverse transcription PCR (RT-PCR) has become an important tool for quantifying changes in messenger RNA (mRNA) populations following differential development or stimulation of tissues or cells. However, the best methods for conducting such experiments and analyzing the resultant data remain an issue of discussion. In this report we describe an appropriate experimental methodology and the computer programs necessary to generate a meaningful statistical analysis of the combined biological and experimental variability in such experiments. Specifically, logarithmic transformations of raw fluorescence data from the log-linear portion of real-time PCR growth curves for both target and reference genes are analyzed using a SAS/STAT Mixed Procedure program specifically designed to give a point estimate of the relative expression ratio of the target gene with associated 95% confidence interval. The program code is open-source and is printed in the text.

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Algorithms Animals Caloric Restriction Gene Expression Profiling Liver Mice Online Systems Programming Languages Reproducibility of Results Reverse Transcriptase Polymerase Chain Reaction Sensitivity and Specificity Software User-Computer Interface Evaluation Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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Peyton Cook, Chunxiao Fu, Morgen Hickey, Eun-Soo Han, Kenton S Miller. 2004. SAS programs for real-time RT-PCR having multiple independent samples.. https://doi.org/10.2144/04376bin02

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