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STAMP: Extensions to the STADEN sequence analysis package for high throughput interactive microsatellite marker design

2009; BioMed Central; Volume: 10; Issue: 1 Linguagem: Inglês

10.1186/1471-2105-10-41

1471-2105

Lars Kraemer, Bánk Beszteri, Steffi Gäbler‐Schwarz, Christoph Held, Florian Leese, Christoph Mayer, Kevin Pöhlmann, Stephan Frickenhaus,

Genomics and Phylogenetic Studies

Abstract Background Microsatellites (MSs) are DNA markers with high analytical power, which are widely used in population genetics, genetic mapping, and forensic studies. Currently available software solutions for high-throughput MS design (i) have shortcomings in detecting and distinguishing imperfect and perfect MSs, (ii) lack often necessary interactive design steps, and (iii) do not allow for the development of primers for multiplex amplifications. We present a set of new tools implemented as extensions to the S TADEN package, which provides the backbone functionality for flexible sequence analysis workflows. The possibility to assemble overlapping reads into unique contigs (provided by the base functionality of the S TADEN package) is important to avoid developing redundant markers, a feature missing from most other similar tools. Results Our extensions to the S TADEN package provide the following functionality to facilitate microsatellite (and also minisatellite) marker design: The new modules (i) integrate the state-of-the-art tandem repeat detection and analysis software P HOBOS into workflows, (ii) provide two separate repeat detection steps – with different search criteria – one for masking repetitive regions during assembly of sequencing reads and the other for designing repeat-flanking primers for MS candidate loci, (iii) incorporate the widely used primer design program P RIMER 3 into S TADEN workflows, enabling the interactive design and visualization of flanking primers for microsatellites, and (iv) provide the functionality to find optimal locus- and primer pair combinations for multiplex primer design. Furthermore, our extensions include a module for storing analysis results in an SQLite database, providing a transparent solution for data access from within as well as from outside of the S TADEN Package. Conclusion The S TADEN package is enhanced by our modules into a highly flexible, high-throughput, interactive tool for conventional and multiplex microsatellite marker design. It gives the user detailed control over the workflow, enabling flexible combinations of manual and automated analysis steps. The software is available under the OpenBSD License [1, 2]. The high efficiency of our automated marker design workflow has been confirmed in three microsatellite development projects.