Submitted to: Animal Genetics
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/17/1996
Publication Date: N/A
Interpretive Summary: Differences in DNA have become an increasingly useful tool in solving disputed parentage of humans and animals. A large number of genetic markers are now characterized and mapped for cattle. Development of a highly informative set of markers with easily differentiated gene forms is critical for using DNA markers as a means of parentage control. A rapid and efficient nonorganic method was developed to extract DNA from dairy and beef bull semen. Multixplex polymerase chain reaction allows several microsatellite marker systems to be analyzed together, which increases the efficiency and effectiveness of using microsatellites for parentage testing and linkage mapping. A total of 22 microsatellite markers were studied, and six multiplexes were developed for semiautomated fluorescence genotyping and evaluated for parentage testing. The 22 markers excluded at least 99.86% of nonparents when genotypes were known for only an alleged parent and an offspring and at least 99.999% if the genotype of a confirmed parent also was known. In addition to providing a rapid and powerful method for parentage testing in cattle, multiplexes will be useful tools for researchers in forensics, gene mapping, and genome scanning.
Technical Abstract: Six multiplexes developed for semiautomated fluorescence genotyping were evaluated for parentage testing. These multiplexes contained primer pairs for the amplification of 22 microsatellites on 17 bovine autosomes. Exclusion probabilities were determined from genotypes of 1,022 Holstein cattle and 311 beef cattle belonging to five breeds. Two cases were considered: 1) genotypes known for an alleged parent and an offspring but unknown for a confirmed parent and 2) genotypes known for an alleged parent, a confirmed parent, and an offspring. If the alleged parent is not the true parent, then the 22 markers will exclude the alleged parent with a probability of >.9986 for case 1 and with a probability of >.99999 for case 2. On the basis of these exclusion probabilities, the probability that an alleged parent will be falsely included as parent is in the range of 1/716 to 1/2845 for case 1 and 1/1.2 million to 1/159,753 for case 2. In addition to these results, a rapid and efficient nonorganic method for extraction of DNA from semen was developed.