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Rapid and efficient screening of a Representational Difference Analysis library using reverse Southern hybridisation: Identification of genetic differences between Haemophilus parasuis isolates

Lancashire, J.F. and Turni, C. and Blackall, P.J. and Jennings, M.P. (2007) Rapid and efficient screening of a Representational Difference Analysis library using reverse Southern hybridisation: Identification of genetic differences between Haemophilus parasuis isolates. Journal of Microbiological Methods, 68 . pp. 326-330.

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Article Link(s): http://dx.doi.org/10.1016/j.mimet.2006.09.011

Publisher URL: http://www.elsevier.com

Abstract

Representational Difference Analysis (RDA) is an established technique used for isolation of specific genetic differences between or within bacterial species. This method was used to investigate the genetic basis of serovar-specificity and the relationship between serovar and virulence in Haemophilus parasuis. An RDA clone library of 96 isolates was constructed using H. parasuis strains H425(P) (serovar 12) and HS1967 (serovar 4). To screen such a large clone library to determine which clones are strain-specific would typically involved separately labelling each clone for use in Southern hybridisation against genomic DNA from each of the strains. In this study, a novel application of reverse Southern hybridisation was used to screen the RDA library: genomic DNA from each strain was labelled and used to probe the library to identify strain-specific clones. This novel approach represents a significant improvement in methodology that is rapid and efficient.

Item Type:Article
Corporate Creators:Animal Science
Additional Information:© Elsevier.
Keywords:Representational difference analysis (RDA); southern hybridisation; Haemophilus parasuis.
Subjects:Veterinary medicine > Diseases of special classes of animals > Swine
Science > Biology > Genetics
Veterinary medicine > Veterinary bacteriology
Deposited On:18 Mar 2008
Last Modified:07 Jun 2011 23:23

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