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Dual-genome primer design for construction of DNA microarrays
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Evolution, Genomics and Systematics, Molecular Evolution.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Evolution, Genomics and Systematics, Molecular Evolution.
2005 (Swedish)In: Bioinformatics, ISSN 1367-4803, E-ISSN 1367-4811, Vol. 21, no 3, 325-332 p.Article in journal (Refereed) Published
Abstract [en]

Motivation: Microarray experiments using probes covering a whole transcriptome are expensive to initiate, and a major part of the costs derives from synthesizing gene-specific PCR primers or hybridization probes. The high costs may force researchers to limit their studies to a single organism, although comparing gene expression in different species would yield valuable information.

Results: We have developed a method, implemented in the software DualPrime, that reduces the number of primers required to amplify the genes of two different genomes. The software identifies regions of high sequence similarity, and from these regions selects PCR primers shared between the genomes, such that either one or, preferentially, both primers in a given PCR can be used for amplification from both genomes. To assure high microarray probe specificity, the software selects primer pairs that generate products of low sequence similarity to other genes within the same genome. We used the software to design PCR primers for 2182 and 1960 genes from the hyperthermophilic archaea Sulfolobus solfataricus and Sulfolobus acidocaldarius, respectively. Primer pairs were shared among 705 pairs of genes, and single primers were shared among 1184 pairs of genes, resulting in a saving of 31% compared to using only unique primers. We also present an alternative primer design method, in which each gene shares primers with two different genes of the other genome, enabling further savings.

Place, publisher, year, edition, pages
2005. Vol. 21, no 3, 325-332 p.
Keyword [en]
DualPrime, Primer, Microarrays
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-76780DOI: 10.1093/bioinformatics/bti001OAI: oai:DiVA.org:uu-76780DiVA: diva2:104692
Available from: 2006-03-08 Created: 2006-03-08 Last updated: 2017-12-14Bibliographically approved

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Bernander, Rolf

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