Fast establishment of single-strand conformation polymorphism by targeted primer development in Pityogenes chalcographus (Coleoptera, Scolytinae)


Arthofer W., Avtzis D.N., and Stauffer C.


Single strand conformation polymorphism (SSCP) is a widespread method for mutation detection in biomedical research. Yet, its potential as a tool for population genetics is still not fully utilized. Based on mitochondrial DNA sequences of 96 specimens of the wood boring beetle Pityogenes chalcographus we constructed a phylogenetic tree of European populations. This tree consisted of six broadly sympatric diverged lineages containing in total 34 haplotypes. Genetic regions of high mutational activity were determined and used for targeted SSCP primer development. In an SSCP mass screening of 427 individuals more than 80% could be assigned to a distinct clade, revealing the insect's genetic structure in Europe. It was demonstrated that analysis of known sequences allows the setup of a functional SSCP protocol within less than two weeks of working time and that phylogenetic data may be retrieved with high accuracy and significantly reduced costs compared to direct sequencing of PCR products.



Electrophoresis 28, 1046-1052.




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