Molecular and genetic studies of
canine disease phenotypes can be limited by the amount of
DNA available for analysis. New methods have been developed to amplify the genomic
DNA of a species producing large quantities of
DNA from small starting amounts. Whole genome amplification (WGA) of
DNA is now being used in human studies, although this technique has not been applied extensively in veterinary research. We evaluated WGA of canine
DNA for suitability in a range of molecular tests.
DNA from 93 canine blood extracted and 18 buccal swab samples was subjected to WGA using the GenomiPhi kit (Amersham). Genomic
DNA was compared with WGA product using a range of techniques, including reference strand-mediated conformation analysis, denaturing high-performance liquid chromatography analysis, microsatellite genotyping, direct
DNA sequencing, and single nucleotide polymorphism allelic discrimination. All samples amplified well, giving an average yield of 3 mug of
DNA from 2.5 ng of starting material. Extremely high levels of experimental reproducibility and concordance were observed between source and WGA
DNA samples for all analyses used: greater than 95% for blood extracted
DNA and greater than 80% for buccal swab
DNA. These studies clearly demonstrate the usefulness of WGA of canine
DNA as a means of increasing
DNA quantities for canine studies. This technique will have major implications for future veterinary research.