Technical note: Whole-genome amplification of DNA extracted from cattle semen samples.
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 microg 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.
Hawken
R J
RJ
CSIRO Livestock Industries, Queensland Biosciences Precinct, St Lucia, 4067, Queensland, Australia. Rachel.Hawken@csiro.au
Cavanagh
J A L
JA
Meadows
J R S
JR
Khatkar
M S
MS
Husaini
Y
Y
Zenger
K R
KR
McClintock
S
S
McClintock
A E
AE
Raadsma
H W
HW
D000818:Animals; D002417:Cattle / Q000235:genetics*; D004247:DNA / Q000032:analysis* / Q000737:chemistry; D016678:Genome / Q000235:genetics*; D005838:Genotype; D008297:Male; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D040641:Quantitative Trait Loci; D011933:Reagent Kits, Diagnostic; D012661:Semen / Q000737:chemistry*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment
D011933:Reagent Kits, Diagnostic; D004247:DNA