FULL LENGTH REPRESENTATIONAL CDNA LIBRARY ENRICHMENT

描述：(申请人描述) 每三个美国人中就有一个会患上癌症。连 尽管研究和医学的进步给了我们更大的 对癌症的理解也揭示了这一问题的巨大复杂性 疾病。加深对肿瘤及其形成的认识 肿瘤组织，有兴趣识别所有 肿瘤细胞表达的遗传信息，并将其与 在正常细胞中表达遗传信息。此应用程序描述 为开发创新技术而进行的拟议研究 高效生成具有代表性的全长cDNA文库 这些文库的标准化和减去富集化。饱满 长度cDNA是那些包含整个mrna拷贝的克隆。 它们的派生序列。代表性文库是 文库中存在的每个信使核糖核酸物种至少包含一个cdna。 起始组织。在典型细胞中的mRNA中，普遍存在的和 中频类占总信使核糖核酸的50%到65% 质量，而只代表大约1000到2000个不同的mRNA。冗余 这两个频率类别的mRNAs的识别注定要成为 在一个随机的cDNA发现程序中压倒性地。因此，它是非常有用的 能够构建归一化文库和消减文库。规格化 库是包含每个库的近似摩尔副本的库 RNA来源的mRNA转录本。减去的文库是指 包含代表两个RNA来源之间差异的克隆。这个 这些技术的发展将为许多 他们的目标是进一步了解 肿瘤的产生和治疗方法的发展 这些疾病。

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.