GENETICS AND GENOME BANKING CORE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 导言：CORE的工作重点是遗传资源库和微卫星的产生，以分配亲子关系和确定等位基因频率。 方法：常规采集血液，扩增小串联重复序列(STR)。扩增产物被送往加州大学戴维斯分校进行大小测定。测序仪输出被发回给我们，Core工作人员对每个等位基因进行最终大小解释并指定基因类型。结果/讨论：在过去的一年里，我们收到了594份来自1983年至2007年出生的动物的STR档案，这些档案基于一个由13个基因座组成的较老的STR小组。根据这些，我们计算了等位基因频率，并确定了当比较2000年前和2000年后出生的动物时，杂合性水平没有改变。我们已经使用230只动物的34个基因座的扩展面板建立了STR图谱(另有190只动物等待加州大学戴维斯分校的片段大小测定)，其中包括216名印度人和14名中国人。对这些STR的分析同样显示出高度的杂合性，并不显著偏离平衡条件下的预期，例如完全随机交配。(观察杂合度为0.75，预期杂合度为0.77)。在216只印度动物中，有22只后代，109只潜在的母马和44只潜在的父系。我们已经确定了其中11只动物的父亲身份和4只动物的母亲身份。我们还没有所有父母的STR档案，当这些档案到达时，最终的亲子鉴定将会完成。我们采集了817只动物的血液样本，并获得了362只动物的DNA提取液。至此，已建立并冷冻保存了100个成纤维细胞系，另有50个细胞系正在培养中。

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.