Czech J. Anim. Sci., 2005, 50(11):499-502 | DOI: 10.17221/4254-CJAS
Implications for the use of horse hair roots as a DNA source for microsatellite typing
- Immuno- and Cytogenetics Department, National Research Institute of Animal Production, Balice, Poland
Hair roots are a very attractive source of DNA for microsatellite-based parentage control of breeding animals. However, unlike blood samples, irregular DNA typing results have been observed in assays utilizing hair follicles. The amount of starting material and DNA preparation method are the crucial factors. In order to improve DNA typing results for horse hair roots, two quick preparation methods and additional purification steps were evaluated. PCR efficiency for each approach was expressed as percentage of samples with complete DNA profiles for 12 horse microsatellites. The lowest percentage (22%) of complete DNA profiles was obtained for samples prepared by the proteinase K digestion method. The best genotyping results (94%) were achieved after phenol-chloroform extraction of DNA from samples prepared by the proteinase K digestion method. Direct cleanup of DNA samples with an ethanol-sodium acetate mixture gave comparably good results of microsatellite genotyping (91%). DNA preparation from hair roots with proteinase K digestion followed by DNA purification with ethanol was chosen as the most efficient approach for horse DNA typing under parentage testing.
Keywords: hair follicles; short tandem repeats; multiplex PCR
Published: November 30, 2005 Show citation
References
- Drissing J., Rudbeck L., Marcher H. (1996): A five minute procedure for extraction of genome DNA from whole blood, semen and forensic stain for PCR. In: Carracedo A. (ed.): Advances in Forensic Haemogenetics. SpringerVerlag, New York. 269-271.
Go to original source... - Ellegren H., Johansson M., Sandberg K., Andersson L. (1992): Cloning of highly polymorphic microsatellites in the horse. Anim Genet., 23, 132-133. Go to original source...
Go to PubMed... - Healy P.J., Dennis J.A., Moule J.F. (1995): Use of hair root as a source of DNA for the detection of heterozygotes for recessive defects in cattle. Aust. Vet. J., 72, 392. Go to original source... Go to PubMed...
- Kalmar T., Bachrati C.Z., Marcsik A., Rasko I. (2000): A simple and efficient method for PCR amplifiable DNA extraction from ancient bones. Nucleic Acids Res., 28, E67-e67. Go to original source... Go to PubMed...
- Kawasaki E.S. (1990): Sample preparation from blood, cells and other fluids. In: Innis M.A., Gelfand D.H., Sninsky J.J., White T.J. (eds.): PCR Protocols: a Guide to Methods and Applications. Academic Press, New York. 146-152. Go to original source...
- Kochl S., Niederstatter H., Parson W. (2005): DNA extraction and quantitation of forensic samples using the phenol-chloroform method and real-time PCR. Meth. Mol. Biol., 297,13-30.
- Sambrook J., Fritsch E.F., Maniatis T. (1989): Molecular Cloning: A Laboratory Manual. 2 nd ed. Cold Spring Harbor Laboratory, New York, Appendix E. 3-5.
- StockMarks® for Horse and Cattle Paternity PCR Typing Kits (2002): Protocol, Applied Biosystems, Foster City, California (USA), Revision C. 14.
This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY NC 4.0), which permits non-comercial use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.