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3PD: Rapid design of optimal primers for chromosome conformation capture assays

Sebastian Fröhler and Christoph Dieterich*

Author Affiliations

Berlin Institute for Medical Systems Biology at the Max Delbrück Center for Molecular Medicine, Robert-Rössle-Strasse 10, 13125 Berlin-Buch, Germany

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BMC Genomics 2009, 10:635  doi:10.1186/1471-2164-10-635

Published: 29 December 2009

Abstract

Background

Higher eukaryotes control the expression of their genes by mechanisms that we are just beginning to understand. A complex layer of control is the dynamic spatial organization of the nucleus.

Results

We present a bioinformatics solution (3PD) to support the experimentalist in detecting long-ranging intra or inter chromosomal contacts by Chromosome conformation capture (3C) assays. 3C assays take a snapshot of chromosomal contacts by a fixation step and quantify them by PCR. Our contribution is to rapidly design an optimal primer set for the crucial PCR step. Our primer design reduces the level of experimental error as primers are highly similar in terms of physical properties and amplicon length. All 3C primers are compatible with multiplex PCR reactions. Primer uniqueness is checked genome-wide with a suitable index structure.

Conclusions

In summary, our software 3PD facilitates genome-wide primer design for 3C experiments in a matter of seconds. Our software is available as a web server at: http://www.pristionchus.org/3CPrimerDesign/ webcite.