Cloning of 400 full-length large cDNAs

描述(申请人提供)：最新的人类基因组“构建34”包含21,335个独特的参考序列，代表17,869个独特的基因座。对开放阅读框架(ORF)大小的调查显示，635个参考序列含有大于5kb的ORF，但其中只有11个较大的克隆可通过哺乳动物基因集(MGC)进行商业使用。获得感兴趣基因的物理克隆对于功能分析是绝对必要的。Origene Technologies，Inc.开发了20个最先进的cDNA库，即快速筛选阵列式cDNA库面板，以支持高通量全长cDNA克隆。到目前为止，我们还从我们的初级cDNA文库(非排列的)中分离并测序了800,000个个体克隆，并发布了20,000个独特的全长基因，这是业内最大的，用于功能基因组研究。该项目的第一阶段旨在优化实验，以获得400个非常大的转录本，包括鉴定与每个基因相关的部分重叠克隆。这将需要使用我们的快速筛选技术优化用于克隆的引物设计。该项目的第二阶段将侧重于对其中400个长克隆进行克隆和完整测序，以便向研究界提供这些克隆。我们相信我们的技术平台是可靠的和创新的，这个项目的结果肯定会取得商业成功。

DESCRIPTION (provided by applicant): The latest build of human genome "build 34" contains 21,335 unique reference sequences that represent 17,869 unique loci. A survey of the open reading frame (ORF) size reveals that 635 reference sequences contain ORFs greater than 5 kb, but only 11 of those larger clones are commercially available through the Mammalian Gene Collection (MGC). The availability of a physical clone of the gene of interest is absolutely essential for functional analyses. OriGene Technologies, Inc. has developed 20 state-of-the-art cDNA libraries, the Rapid-Screen Arrayed cDNA library Panels, to support high-throughput full-length cDNA cloning. To date, we have also isolated and sequenced 800,000 individual clones from our primary cDNA libraries (non-arrayed) and have released 20,000 unique full-length genes, the largest in the industry, for functional genomics research. Phase I of this project aims to optimize experiments in order to obtain 400 of those very large transcripts, including the identification of partial, overlapping clones associated with each gene. This will require the optimization of primer design for cloning using our Rapid-Screen technology. Phase II of the project will focus on the cloning and complete sequencing of 400 of those long clones to make them available to the research community. We believe that our technology platform is reliable and innovative, and the outcome of this project will be assuredly a commercial success.