FULL LENGTH REPRESENTATIONAL CDNA LIBRARY ENRICHMENT

全长代表性 CDNA 文库富集

• 批准号: 6072254
• 负责人: Frederick S Hagen
• 金额: $ 0.15万
• 依托单位: ICOGENEX CORPORATION
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-09-30 至 1999-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6072254
• 关键词: DNA replication; Escherichia coli; bacterial genetics; complementary DNA; crosslink; genetic library; messenger RNA; method development; molecular cloning; neoplasm /cancer genetics; nucleic acid chemical synthesis; plasmids; polymerase chain reaction; southern blotting; subtraction hybridization

DESCRIPTION: (Applicant's Description) Cancer will affect one out of every three Americans in their lifetime. Even though advances in research and medicine have given us a greater understanding of cancer it has also revealed the great complexity of this disease. Towards a greater understanding of neoplasms and the formation of neoplastic tissue, there is an interest in the identification of all expressed genetic information of neoplastic cells and to compare this to expressed genetic information in normal cells. This application describes proposed research for the development of innovative technologies for the efficient generation of representational full length cDNA libraries and for the normalization and subtractive enrichment of these libraries. Full Length cDNAs are those clones that contain a copy of the entire mRNA sequence from which they were derived. Representational libraries are libraries that contain at least one cDNA for every mRNA species present in the starting tissue. Of the mRNAs in a typical cell, the prevalent and intermediate frequency classes comprise between 50 to 65 % of the total mRNA mass while representing only about 1000 to 2000 different mRNAs. Redundant identification of mRNAs of these two frequency classes is destined to become overwhelming in a random cDNA discovery program. Thus it is very useful to be able to construct normalized and subtracted libraries. Normalized libraries are libraries which contain a near molar equivalent copy of each mRNA transcript of the RNA source. Subtracted libraries are libraries which contain clones representing the difference between two sources of RNA. The development of these technologies will provide tools for the many investigators who have as their goal the further understanding of the generation of neoplasms and the development of methods for the treatment of these diseases.

描述：（申请人的描述） 癌症将影响每三个美国人中的一个。 甚至 尽管研究和医学的进步给了我们更大的 它也揭示了癌症的巨大复杂性， 疾病 为了更好地了解肿瘤和形成 肿瘤组织，有兴趣在所有的识别 表达肿瘤细胞的遗传信息，并将其与 在正常细胞中表达遗传信息。 本申请描述 为开发创新技术而进行的拟议研究， 有效产生代表性全长cDNA文库， 这些文库的标准化和消减富集。 充分 长度cDNA是那些包含完整mRNA拷贝的克隆 它们所衍生的序列。 代表性库是 文库包含存在于细胞中的每个mRNA种类的至少一种cDNA， 起始组织。 在典型细胞中的mRNA中， 中间频率类占总mRNA的50 - 65 质量，而仅代表约1000至2000种不同的mRNA。 冗余 这两种频率类别的mRNA的鉴定注定要成为 在随机cDNA发现计划中占压倒性优势。 因此， 能够构建标准化和消减文库。 归一化 文库是含有每一种的接近摩尔当量拷贝的文库 RNA来源的mRNA转录本。 子库是指 包含代表两种RNA来源之间差异的克隆。 的 这些技术的发展将为许多人提供工具， 研究人员的目标是进一步了解 肿瘤的产生和肿瘤治疗方法的发展 这些疾病。