ONCOGENE ANALYSIS FOR EPIDEMIOLOGIC STUDIES

合同的主要目标是提供开发和 对收集的人体样本进行分子分析， 流行病学研究。 这些研究涉及是否存在 肿瘤中不同的DNA损伤模式由不同的 致癌物，以及基因-环境相互作用在 疾病 虽然没有一项研究将涉及所有测定， 承包商将与各种分子 技术. 基本技术包括： 1.从冷冻的肿瘤组织中提取高分子量DNA， 来自外周白色细胞的冷冻样品，以及来自切片机 固定组织切片，沿着进行组织学分析， 肿瘤样品的显微切割。 2. PCR扩增选定的基因，例如，第53页。 能力 从显微切割的固定组织样品中扩增DNA， 尤其重要。 3.单链构象多态性（SSCP）分析 扩增DNA以筛选突变。 4. PCR扩增DNA的直接测序 足以将10个细胞中的1个中存在的突变体鉴定为 最低标准。 5.与正常相比，肿瘤中杂合性缺失的测定 组织样本 6.已知等位基因变体的PCR-RFLP分析。

The primary goal of the contract is to provide development and application of molecular assays on human specimens gathered in epidemiologic studies. These studies address whether there are different patterns of DNA damage in tumors initiated by different carcinogens, and the role of gene-environment interaction in disease. Although no one study will involve all assays, the contractor will be working with a variety of molecular techniques. Basic techniques include the following: 1. Extract high molecular weight DNA from frozen tumor tissue and from frozen samples of peripheral white cells, and from microtome slices of fixed tissues, along with histologic analysis and microdissection of tumor samples. 2. PCR amplification of selected genes, e.g., p53. Ability to amplify DNA from microdissected fixed tissue samples is particularly important. 3. Single strand conformation polymorphism (SSCP) analysis of amplified DNA to screen for mutations. 4. Direct sequencing fo PCR amplified DNA with sensitivity adequate to identify mutants present in 1 of 10 cells as a standard minimum. 5. Assay for loss of hetrozygosity in tumor compared to normal tissue samples. 6. PCR-RFLP analysis of known allelic variants of genes.