GENETICS AND GENOME BANKING CORE

遗传学和基因组库核心

• 批准号: 8173011
• 负责人: Hans Michael Kubisch
• 金额: $ 6.18万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8173011
• 关键词: Alleles; Animals; Back; Blood; Blood Banks; Blood specimen; Cell Line; Chinese People; Computer Retrieval of Information on Scientific Projects Database; DNA; Equilibrium; Equipment and supply inventories; Fibroblasts; Funding; Gene Frequency; Generations; Genetic; Genome; Genotype; Grant; Institution; Methods; Microsatellite Repeats; Output; Parents; Partner in relationship; Research; Research Personnel; Resources; Source; Tandem Repeat Sequences; United States National Institutes of Health; base; father role; genetic resource; male; offspring

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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 中心，不一定是研究者的机构。 简介：核心的工作重点是遗传资源库和微卫星的生成，以分配亲子关系和确定等位基因频率。 方法：在常规清查期间收集血液，我们从中扩增了小串联重复序列（STR）。 PCR 产物被送往加州大学戴维斯分校进行大小测定。测序仪的输出被发送回给我们，核心工作人员对每个等位基因进行最终的大小解释并指定基因型。 结果/讨论：在过去的一年中，我们收到了 1983 年至 2007 年出生的动物的 594 个 STR 谱，这些谱基于包含 13 个基因座的旧 STR 组。由此我们计算了等位基因频率，并确定当比较 2000 年之前和之后出生的动物时，杂合性水平没有变化。我们使用来自 230 只动物的 34 个基因座的扩展组建立了 STR 谱（另外 190 只动物在加州大学戴维斯分校等待片段大小测定），其中包括 216 只印度雄性和 14 只中国雄性。 对这些 STR 的分析同样显示出高度的杂合性，与平衡条件下（例如完全随机交配）下的预期并没有显着偏离。 （观察杂合度：0.75，预期杂合度：0.77）。 216 头印度动物中有 22 只后代、109 只潜在母猫和 44 只潜在父猫。我们已为其中 11 只动物确定了亲子关系，并为其中 4 只动物确定了母子关系。我们尚未将所有父母的 STR 资料存档，最终的亲子关系将在这些资料送达后进行。 我们采集了 817 只动物的血液样本，并获得了 362 只动物的 DNA 提取物。这使得血库动物数量达到 4,356 只。总共建立并冷冻保存了 100 个成纤维细胞系，另外 50 个细胞系目前正在培养中。