REST 2009

version 1  released  22. December 2009

REST 2009 Software is a standalone software tool, developed by M. Pfaffl (Technical University Munich) and QIAGEN, for analysis of gene expression data from quantitative real-time PCR experiments. Click the "Resources" tab to download REST 2009 Software free of charge.


Download newest version REST 2009  =>  http://www.REST.de.com
Qiagen download page  via  Analysis Software


Download User Guide  =>  REST 2009 Software User Guide - English (PDF)


REST 2009 Software
For gene expression analysis using real-time PCR data from the Rotor-Gene Q and other cyclers
  • Estimates up and down regulation for gene expression studies
  • Analysis using randomization and bootstrapping techniques
  • Graphical data output via whisker-box plots
Principle
REST 2009 Software applies a mathematic model that takes into account the different PCR efficiencies of the gene of interest and reference genes. Compared to using a single reference gene, using multiple reference genes for normalization can improve the reliability of results.

Traditional relative quantitation enables estimation of gene expression. However, this method does not provide statistical information that is suitable for comparing expression in groups of treated and untreated samples in a robust manner. The integrated randomization and bootstrapping methods used in REST 2009 Software test the statistical significance of calculated expression ratios and can be used even when the data includes outliers. REST 2009 Software provides additional features for convenient and robust data analysis (see table "Convenient and robust data analysis").


Convenient and robust data analysis


Feature Description
REST RG mode An optional input method allows users to copy and paste results from a Rotor-Gene Q comparative quantitation analysis rather than importing standard curve and CT results.
Whisker-box plots export Expression variation for each gene is visualized in a whisker-box plot to highlight potential issues, such as a distribution skew. Whisker-box plots are exported by right-clicking the graph.
Improved randomization Randomization algorithms have been improved for better confidence intervals and more accurate p values.
Handling of standard curve variation Improvements have been made to the calculation of confidence intervals and p values. Efficiency is determined using the best fit for the standard curve and is used in the randomization process.


Applications REST 2009 Software is suitable for gene expression analysis using real-time PCR data from the Rotor-Gene Q and other cyclers.


Cited references

  1. Pfaffl, M.W. (2001) A new mathematical model for relative quantification in real-time RT-PCR. NAR 29, e45
  2. Pfaffl, M.W., Horgan, G.W., Dempfle, L. (2002) Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. NAR 30, e36
  3. Vandesompele J. et. al. (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 3, research0034.1
  4. Relative Expression Software Tool http://rest.gene-quantification.info
  5. Randomization Tests http://ordination.okstate.edu/permute.htm
  6. Introduction to Randomization Tests http://www.bioss.ac.uk/smart/unix/mrandt/slides/frames.htm
  7. Resampling Statistics: Randomization and the Bootstrap http://www.uvm.edu/~dhowell/StatPages/Resampling/Resampling.html
  8. Quartiles and Box and Whisker Plots http://www.regentsprep.org/regents/math/algebra/AD3/boxwhisk.htm


REST 2009 Software is intended for molecular biology applications. This software is neither intended for the diagnosis, prevention, or treatment of a disease, nor has it been validated for such use either alone or in combination with other products. Therefore, the performance characteristics of the product for clinical use (i.e., diagnostic, prognostic, therapeutic, or blood banking) are unknown.


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