Multiple Displacement Amplification for Generating an Unlimited Source of DNA for Genotyping in Nonhuman Primate Species |
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| Authors: | A.-C. Rönn O. Andrés M. W. Bruford B. Crouau-Roy G. Doxiadis X. Domingo-Roura A. D. Roeder E. Verschoor H. Zischler A.-C. Syvänen |
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| Affiliation: | (1) Molecular Medicine, Department of Medical Sciences, Uppsala University, Uppsala, Sweden;(2) Genètica de la Conservació, Institut de Recerca i Tecnologia Agroalimentàries, Centre de Cabrils, Cabrils, Spain;(3) Cardiff School of Biosciences, Cardiff University, Cardiff, UK;(4) Evolution et Diversité Biologique UMR 5174, Université Paul Sabatier Toulouse, Toulouse, France;(5) Department of Comparative Genetics and Refinement, Biomedical Primate Research Centre, Rijswijk, The Netherlands;(6) Department of Virology, Biomedical Primate Research Centre, Rijswijk, The Netherlands;(7) Institute of Anthropology, Johannes-Gutenberg University, Mainz, Germany |
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| Abstract: | We evaluated a whole genome amplification method—multiple displacement amplification (MDA)—as a means to conserve valuable nonhuman primate samples. We tested 148 samples from a variety of species and sample sources, including blood, tissue, cell-lines, plucked hair and noninvasively collected semen. To evaluate genotyping success and accuracy of MDA, we used routine genotyping methods, including short tandem repeat (STR) analysis, denaturing gradient gel electrophoresis (DGGE), Alu repeat analysis, direct sequencing, and nucleotide detection by tag-array minisequencing. We compared genotyping results from MDA products to genotypes generated from the original (non-MD amplified) DNA samples. All genotyping methods showed good results with the MDA products as a DNA template, and for some samples MDA improved genotyping success. We show that the MDA procedure has the potential to provide a long-lasting source of DNA for genetic studies, which would be highly valuable for the primate research field, in which genetic resources are limited and for other species in which similar sampling constraints apply. |
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| Keywords: | Alu-SINE minisequencing multiple displacement amplification short tandem repeat single nucleotide polymorphism |
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