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An integrated pipeline for the development of novel panels of mapped microsatellite markers for Leishmania donovani complex, Leishmania braziliensis and Leishmania major

Fakhar, M.; Motazedian, M.H.; Daly, D.; Lowe, C.D.; Noyes, H.A. and Kemp, S.J. (2008) An integrated pipeline for the development of novel panels of mapped microsatellite markers for Leishmania donovani complex, Leishmania braziliensis and Leishmania major. Parasitology, 135 (5). pp. 567-574. ISSN 0031-1820

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Cited 3 times in WoS

Abstract

A panel of microsatellites mapped to the Leishmania genome might make it possible to find associations between specific loci and phenotypic traits. To identify such loci, a Perl programme was written that scans the sequence of a genome and writes all loci containing microsatellites to a MySQL database. The programme was applied to the sequences of the L. braziliensis, L. infantum and L. major genomes. The database is publicly available over the internet: http://www.genomics.liv.ac.uk/tryps/resources.html “Microsatellite Locus Extractor”, and allows the selection of mapped microsatellites that meet user defined criteria from a specified region of the selected genome. The website also incorporates a primer design pipeline that will design primers to amplify the selected loci. Using this pipeline 12 out of 17 primer sets generated polymorphic PCR products. A tailed primer protocol was used to label all microsatellite primers with a single set of labelled primers. To avoid the culture of parasites prior to genotyping, sets of nested PCR primers were developed to amplify parasite DNA eluted from microscope slides. The limit of detection was approximately 1.6 parasite equivalents. However only 6/56 DNA from slides stored at ambient temperature for over six months gave positive PCR results.

Item Type:Article
Additional Information:Published online by Cambridge University Press 27 Mar 2008. Issue 05 - Apr 2008. Citation: M. FAKHAR, M. H. MOTAZEDIAN, D. DALY, C. D. LOWE, S. J. KEMP and H. A. NOYES (2008). An integrated pipeline for the development of novel panels of mapped microsatellite markers for Leishmania donovani complex, Leishmania braziliensis and Leishmania major. Parasitology, 135, pp 567-574 doi:10.1017/S0031182008004186
Uncontrolled Keywords:Leishmania donovani complex; Leishmania braziliensis, Leishmania major; Leishmania infantum; Leishmania Viannia; microsatellite; tailed primers; nested PCR; Iran; VISCERAL LEISHMANIASIS; SOUTHERN IRAN; INFANTUM; POLYMORPHISMS; INCREASE; PRIMERS
Subjects:Q Science > QH Natural history > QH301 Biology
Q Science > QH Natural history > QH426 Genetics
Departments, Research Centres and Related Units:Academic Faculties, Institutes and Research Centres > Faculty of Science > Department of Biological Sciences
DOI:10.1017/S0031182008004186
Publisher's Statement:M. FAKHAR, M. H. MOTAZEDIAN, D. DALY, C. D. LOWE, S. J. KEMP and H. A. NOYES (2008). An integrated pipeline for the development of novel panels of mapped microsatellite markers for Leishmania donovani complex, Leishmania braziliensis and Leishmania major. Parasitology, 135 , pp 567-574 doi:10.1017/S0031182008004186 Copyright © Cambridge University Press 2008 http://journals.cambridge.org/action/displayJournal?jid=PAR
Refereed:Yes
Status:Published
ID Code:749
Deposited On:20 Aug 2008 14:12
Last Modified:06 Feb 2012 17:45

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