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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.
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Download
newest
version REST 2009 =>
http://www.REST.de.com
Qiagen
download page via Analysis
Software
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| Download
User Guide => REST
2009 Software User Guide - English (PDF) |
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| 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
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| 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"). |
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| Convenient and robust data
analysis |
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| Feature
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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.
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| 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.
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| Applications |
REST 2009
Software is suitable for gene expression
analysis using real-time PCR data from the Rotor-Gene Q and other
cyclers. |
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| Cited
references |
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- Pfaffl, M.W.
(2001) A new mathematical model for relative quantification in
real-time RT-PCR. NAR 29, e45
- 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
- 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
- Relative Expression Software Tool http://rest.gene-quantification.info
- Randomization Tests http://ordination.okstate.edu/permute.htm
- Introduction to Randomization Tests http://www.bioss.ac.uk/smart/unix/mrandt/slides/frames.htm
- Resampling Statistics: Randomization and
the Bootstrap http://www.uvm.edu/~dhowell/StatPages/Resampling/Resampling.html
- Quartiles and Box and Whisker Plots http://www.regentsprep.org/regents/math/algebra/AD3/boxwhisk.htm
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| 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. |