Single bovine sperm sex typing by amelogenin nested PCR.
Sex-sorted bovine semen has become a valuable tool in animal production for sex preselection. Development of novel sperm sexing technologies, or evaluation of the quality of existing methods, often requires a single-sperm, sex-typing method that is reliable and easy to perform. In the present study, we report the development, validation, and application of a simple, reliable, and cost-effective method for single-sperm sex typing using nested polymerase chain reaction (PCR), based on the amelogenin gene. Several hundred single sperm were isolated using a simple manual technique, or a high-speed flow-sorter, and were successfully sex-typed using the amelogenin nested PCR. Based on the pooled results of individual sperm, there was no significant difference in the semen sex ratio of unsorted (44.6% X-sperm and 55.4% Y-sperm) or X/Y-sorted semen (91.4% X-sperm and 94.0% Y-sperm), as compared to the expected ratio in unsorted semen or the post-sorting reanalysis data, respectively. The amelogenin single-sperm sexing method was an adaptable, accurate, and reliable tool for single-sperm sex typing.
Colley
A
A
Department of Animal and Poultry Science, University of Guelph, Guelph, ON N1G 2W1, Canada.
Buhr
M
M
Golovan
S P
SP
D053523:Amelogenin / Q000235:genetics*; D000818:Animals; D002417:Cattle / Q000235:genetics* / Q000502:physiology; D002469:Cell Separation / Q000379:methods / Q000662:veterinary; D008297:Male; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000662:veterinary; D012730:Sex Chromosomes; D012732:Sex Determination Analysis / Q000379:methods* / Q000662:veterinary; D013094:Spermatozoa / Q000166:cytology / Q000378:metabolism*
D053523:Amelogenin