Whole-genome amplification of DNA extracted from cattle semen samples

Hawken, R.J., Cavanagh, J.A.L., Meadows, J.R.S., Khatkar, M.S., Husaini, Y., Zenger, K.R., McClintock, S., McClintock, A.E., and Raadsma, H.W. (2006) Whole-genome amplification of DNA extracted from cattle semen samples. Journal of Dairy Science, 89 (6). pp. 2217-2221.

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Abstract

The bovine genome sequence project and the discovery of many thousands of bovine single nucleotide polymorphisms has opened the door for large-scale genotyping studies to identify genes that contribute to economically important traits with relevance to the beef and dairy industries. Large amounts of DNA will be required for these research projects. This study reports the use of the whole-genome amplification (WGA) method to create an unlimited supply of DNA for use in genotyping studies and long-term storage for future gene discovery projects. Two commercial WGA kits (GenomiPhi, Amersham Biosciences, Sydney, Australia, and REPLI-g, Qiagen, Doncaster, Australia) were used to amplify DNA from straws of bull semen, resulting in an average of 7.2 and 67 g of DNA per reaction, respectively. The comparison of 3.5 kb of sequences from the amplified and unamplified DNA indicated no detectable DNA differences. Similarly, gene marker analysis conducted on genomic DNA and DNA after WGA indicated no difference in marker amplification or clarity and accuracy of scoring for approximately 10,000 single nucleotide polymorphism markers when compared with WGA samples genotyped in duplicate. These results illustrate that WGA is a suitable method for the amplification and recovery of DNA from bull semen samples for routine genomic investigations.

Item ID: 9784
Item Type: Article (Refereed Research - C1)
Keywords: whole-genome amplification; GenomiPhi; REPLI-g; DNA extraction; animal production and genomics
ISSN: 1525-3198
Date Deposited: 30 Mar 2010 23:04
FoR Codes: 07 AGRICULTURAL AND VETERINARY SCIENCES > 0702 Animal Production > 070201 Animal Breeding @ 50%
06 BIOLOGICAL SCIENCES > 0604 Genetics > 060408 Genomics @ 50%
SEO Codes: 83 ANIMAL PRODUCTION AND ANIMAL PRIMARY PRODUCTS > 8303 Livestock Raising > 830302 Dairy Cattle @ 40%
97 EXPANDING KNOWLEDGE > 970106 Expanding Knowledge in the Biological Sciences @ 60%
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