NR AOTN
AU Gutscher,M.; Brem,G.
TI Large scale sample collection and DNA preparation system for scrapie-resistance genotyping in sheep
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - DG-52
PT Konferenz-Poster
AB The establisment of breeding programmes to increase the number of sheep that are naturally resistant to Transmissible Spongiform Encephalopathies necessitates the collection of DNA samples of whole populations. Currently blood samples are mainly collected, a cost and time consuming method associated with stress for the animals. To overcome this drawbacks, we have developed an easy-handling and cost-effectiv tissue collection system. The basic principle of the TypiFix system is simultaneous sampling of tissue while placing an ear tag for individual animal identification. The tissue is collected in a sample container, which preserves the DNA upto 3 years automatically for moleculargenetic analysis (failure rate below 0.1 %). The ear-tag and sample container are attached to each other and preprinted with identical identification numbers to exclude confusion of samples during collection. In combination with a new "one-step" nucleic acid purification method based on chromatographic polymer-containing support DNA preparation of a large number of samples can be achieved in a short time. An average yield of DNA of 16,6 ± 6,7 µg per TypiFix sample was reached, which allows the building up of a DNA bank and further moleculargenetic analysis like paternity control or QTL studies. 363 DNA samples were scrapie-resistance genotyped by Pyrosequencing (failure rate 0.27 %). All 5 known PrP alleles of the ovine prionprotein could be analysed. In addition a very rare allel ARK were found.
AD M. Gutscher, G. Brem, Agrobiogen GmbH, Germany
SP englisch
PO Deutschland