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



Fakhar, M, Motazedian, MH, Daly, D, Lowe, CD, Kemp, SJ and Noyes, HA ORCID: 0000-0002-0656-200X
(2008) An integrated pipeline for the development of novel panels of mapped microsatellite markers for <i>Leishmania donovani</i> complex, <i>Leishmania braziliensis</i> and <i>Leishmania major</i>. PARASITOLOGY, 135 (5). pp. 567-574.

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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 designed against the L. infantum genome 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 6 months gave positive PCR results.

Item Type: Article
Additional Information: ## TULIP Type: Articles/Papers (Journal) ##
Uncontrolled Keywords: Leishmania donovani complex, Leishmania braziliensis, Leishmania major, Leishmania infantum, Leishmania (Viannia), microsatellite, tailed primers, nested PCR, Iran
Subjects: ?? QH301 ??
?? QH426 ??
Divisions: Faculty of Health and Life Sciences
Depositing User: Symplectic Admin
Date Deposited: 20 Aug 2008 13:12
Last Modified: 17 Oct 2023 09:06
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
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URI: https://livrepository.liverpool.ac.uk/id/eprint/749