GENETICS AND GENOME BANKING CORE

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Introduction: The efforts of the Core have focused on banking of genetic resources and on generation of microsatellites to assign paternity and determine allele frequencies. Methods: Blood was collected during routine inventories from which we have amplified small tandem repeats (STRs). The PCR products were sent to UC Davis for size determination. The sequencer output was sent back to us and Core staff makes the final size interpretations of each allele and asigns genotypes. Results/Discussion: During the past year we received 594 STR profiles from animals born from 1983 through 2007 that were based on an older STR panel of 13 loci. From these we have calculated allele frequencies and have determined that the level of heterozygosity has not changed when animals born before and after 2000 are compared. We have established STR profiles using the expanded panel of 34 loci from 230 animals (an additional 190 animals await fragment size determinations at UC Davis), which included 216 Indian and 14 Chinese males. Analysis of these STRs similarly showed a high degree of heterozygosity that does not deviate significantly from what would be expected under equilibrium conditions, such as completely random mating. (observed heterozygosity: 0.75, expected heterozygosity: 0.77). Among the 216 Indian animals were 22 offspring, 109 potential dams and 44 potential sires. We have established paternity for 11 and maternity for 4 of these animals. We do not yet have STR profiles on file for all parents and final parentage determinations will be done when those have arrived. We have collected blood samples from 817 animals, and have obtained DNA extracts from 362 animals. This brings the number of blood-banked animals to 4,356.A total of 100 fibroblast cell lines have been established and cryopreserved, with an additional 50 lines currently in culture.