CORE--MACROMOLECULAR ANALYSIS FACILITY

大分子分析设备由三个独立的但 相关实体：DNA测序设施(DSF)，它执行 自动DNA测序，自动微卫星分析，以及单个 链构象多态(SSCP)分析 分析设施(CAF)，帮助调查人员进行计算 大分子结构分析；和核酸核心设施 (NACF)，一个用于标准化提取DNA的中央实验室 转基因小鼠的各种来源和基因分型。这些资源 从血液、肿瘤样本和石蜡包埋或冷冻中制备DNA 切片(在NACF中)；确定DNA序列并执行微卫星 和SSCP分析(在DSF中)。DNA序列由CAF进行分析。这个 DSF为广泛的基础、临床和研究人员提供帮助 以人口为基础的研究涉及增长控制、增长 抑制、对特定细胞系的承诺、细胞黏附和 移动性、转移、血管生成等领域，以及支持 新的肿瘤易感基因的定位和克隆研究 在已知癌症中筛查肿瘤和基因突变 易感基因与肿瘤和细胞突变的筛查 在已知的癌症易感基因和肿瘤筛查中 已知癌症易感基因的结构性突变和 致癌基因。DSF将快速可靠的序列信息提供给 以高效率和高成本效益的方式开展调查。DNA的数量 DSF确定的序列增加了400%，从1992年的约1500个增加到 1996年7月1日至1997年6月1日期间超过6,000人，以及 可读序列从大约300-350个碱基增加到超过650个碱基 样本。为了生成DS序列，DSF目前在ABI 373上使用 配备拉伸配置的DNA测序仪和最近推出的一台 获得了ABI 377型DNA测序仪。该资源为调查人员提供了 聚合酶链式反应(PCR)产生的DNA，可以产生最多的 准确的DNA序列和在引物设计和维护方面的专业知识 严格的质量控制，确保服务的一致性。一大笔钱 该设施的努力致力于确定 使用新的和改进的荧光染料和更高效的DNA测序 酵素。该设施最近开始使用ABI 377 DNA测序仪进行 自动分析聚合酶链式反应产物的大小和数量。分析 检测聚合酶链式反应产物中的SSCP并通过以下方式进行遗传连锁分析 确定含有特定微卫星的大小和数量 扩增产物。377ABI DNA测序仪也用于执行 微卫星分析用于评估体质和肿瘤特异性 基因组不稳定。

The Macromolecular Analysis Facility consists of three separate but related entities: the DNA Sequencing Facility (DSF), which performs automated DNA sequencing, automated microsatellite analysis, and single- strand conformation polymorphism (SSCP) analysis; the Computational Analysis Facility (CAF), which helps investigators with computational analysis of macromolecular structures; and the Nucleic Acid Core Facility (NACF), a centralized laboratory for standardized DNA extraction from various sources and genotyping of transgenic mice. These resources prepared DNA from blood, tumor samples, and paraffin-embedded or frozen sections (in the NACF); determine DNA sequences and perform microsattelite and SSCP analyses (in the DSF). DNA sequences are analyzed by the CAF. The DSF provides help to a wide range of investigators in basic, clinical and population-based research involved in studies of growth control, growth inhibition, commitment to specific cell lineages, cell adhesion and mobility, metastasis, angiogenesis, and other fields, as well as supports studies on the mapping and cloning of new cancer susceptibility genes and screening for tumor and constitutional mutations in known cancer susceptibility genes and screening for tumor and constitutional mutations in known cancer susceptibility genes and screening for tumor and constitutional mutations in known cancer susceptibility genes and oncogenes. The DSF provides fast and reliable sequence information to investigators in an efficient and cost-effective manner. The number of DNA sequences determined by the DSF increased 400% from about 1,500 in 1992 to over 6,000 between July 1, 1996 and June 1, 1997, and the length of readable sequences increased from about 300-350 to more than 650 bases per sample. To generate the DS sequences, the DSF currently uses on ABI 373 DNA sequencer equipped with stretch configuration and one recently acquired ABI 377 DNA sequencer. The resource provides investigators with the polymerase chain reaction (PCR)-generated DNAs, to produce the most accurate DNA sequences and expertise in primer design and maintain rigorous quality control to ensure consistency of service. A substantial effort of the facility is dedicated to determining optimal conditions for DNA sequencings with new and improved fluorescent dyes and more efficient enzymes. The facility recently began to use the ABI 377 DNA sequencer for automated analysis of the sizes and amounts of PCR products. The analysis detects SSCPs in PCR products and allows genetic linkage analysis by determining the sizes and amounts of specific microsatellite-containing PCR products. The 377 ABI DNA sequencer is also used to perform microsatellite analysis to evaluate constitutional and tumor-specific genomic instability.