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Dual-genome primer design for construction of DNA microarrays
KTH, School of Biotechnology (BIO).ORCID iD: 0000-0002-3627-6899
Department of Molecular Evolution, Evolutionary Biology Center, Uppsala University.
KTH, School of Biotechnology (BIO).ORCID iD: 0000-0002-4657-8532
2005 (English)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]
sulfolobus-solfataricus, gene, coexpression, initiation
National Category
Industrial Biotechnology
Identifiers
URN: urn:nbn:se:kth:diva-14479DOI: 10.1093/bioinformatics/bti001ISI: 000226605700007Scopus ID: 2-s2.0-13844264518OAI: oai:DiVA.org:kth-14479DiVA: diva2:332520
Note
QC 20100830Available from: 2010-08-05 Created: 2010-08-05 Last updated: 2017-12-12Bibliographically approved
In thesis
1. Microarray-based investigation of genome and transcriptome organisation in the archaeon sulfolobus
Open this publication in new window or tab >>Microarray-based investigation of genome and transcriptome organisation in the archaeon sulfolobus
2005 (English)Doctoral thesis, comprehensive summary (Other scientific)
Place, publisher, year, edition, pages
Stockholm: KTH, 2005. 47 p.
Keyword
Biotechnology, microarray, Sulfolobus, gene expression, archaea, mRNA decay, genome evolution, Bioteknik
National Category
Industrial Biotechnology
Identifiers
urn:nbn:se:kth:diva-142 (URN)91-7283-979-1 (ISBN)
Public defence
2005-03-11, Sal D1, Lindstedtsvägen 17, Stockholm, 10:00
Opponent
Supervisors
Note
QC 20100830Available from: 2005-03-03 Created: 2005-03-03 Last updated: 2010-08-30Bibliographically approved

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