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Bioinformatics Advance Access originally published online on March 17, 2005
Bioinformatics 2005 21(11):2573-2579; doi:10.1093/bioinformatics/bti393
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© The Author 2005. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions{at}oupjournals.org

Optimization of cDNA-AFLP experiments using genomic sequence data

Teemu Kivioja 1,*,{dagger}, Mikko Arvas 2, Markku Saloheimo 2, Merja Penttilä 2 and Esko Ukkonen 1

1Department of Computer Science, University of Helsinki Helsinki, PO Box 68, FIN-00014, Finland
2VTT Biotechnology Espoo, PO Box 1500, FIN-02044, Finland

*To whom correspondence should be addressed.

Motivation: cDNA amplified fragment length polymorphism (cDNA-AFLP) is one of the few genome-wide level expression profiling methods capable of finding genes that have not yet been cloned or even predicted from sequence but have interesting expression patterns under the studied conditions. In cDNA-AFLP, a complex cDNA mixture is divided into small subsets using restriction enzymes and selective PCR. A large cDNA-AFLP experiment can require a substantial amount of resources, such as hundreds of PCR amplifications and gel electrophoresis runs, followed by manual cutting of a large number of bands from the gels. Our aim was to test whether this workload can be reduced by rational design of the experiment.

Results: We used the available genomic sequence information to optimize cDNA-AFLP experiments beforehand so that as many transcripts as possible could be profiled with a given amount of resources. Optimization of the selection of both restriction enzymes and selective primers for cDNA-AFLP experiments has not been performed previously. The in silico tests performed suggest that substantial amounts of resources can be saved by the optimization of cDNA-AFLP experiments.

Availability: A Perl implementation of the optimization method is available upon request from the authors.

Contact: Teemu.Kivioja{at}vtt.fi


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