Abstract
Application of the amplified fragment length polymorphism (AFLP) technique to genetic mapping studies requires high quality DNA as a template. In the case of Eimeria spp., this has previously been in the form of chromosomal DNA obtained from purified sporozoites recovered from large numbers of oocysts (generally up to 2×108). In order for the AFLP technique to be more easily applied to studies on the genetics of Eimeria maxima, for which only smaller numbers of oocysts are available, a simplified, more efficient method for the recovery of genomic DNA from small numbers of oocysts was developed. Our new method should also be useful for genetic analyses of other coccidial parasites and for the recovery of AFLP-quality DNA from other pathogens.
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Acknowledgements
The authors wish to acknowledge the technical assistance of Fionnadh Carroll, Pat Hesketh and Andrew Archer. The Biotechnology and Biological Sciences Research Council financially supported this study under grant 201/S15343. The experiments performed in this study comply with UK and EU law.
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Blake, D.P., Smith, A.L. & Shirley, M.W. Amplified fragment length polymorphism analyses of Eimeria spp.: an improved process for genetic studies of recombinant parasites. Parasitol Res 90, 473–475 (2003). https://doi.org/10.1007/s00436-003-0890-x
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DOI: https://doi.org/10.1007/s00436-003-0890-x