END SEQUENCING AND FINGERPRINTING OF MOUSE BAC LIBRARIES

小鼠 BAC 文库的末端测序和指纹图谱

• 批准号: 6451359
• 负责人: SHAYING ZHAO
• 金额: $ 145.83万
• 依托单位: INSTITUTE FOR GENOMIC RESEARCH
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6451359
• 关键词: animal data; animal genetic material tag; animal tissue; artificial chromosomes; genetic library; laboratory mouse; nucleic acid sequence; restriction fragment length polymorphism

As the human project shifts into production sequencing, a plan is developing for sequencing the mouse genome. The current plan employs a "sequence first, map second" strategy which, like the human project, is based on shotgun sequencing of BAC clones comprising contiguous regions of the genome of the selected mouse strain (C57BL6/J). Libraries constructed in bacterial artificial chromosomes (BAC) vectors have become the choice for high throughput genomic sequencing projects because of their cloning capacity and high stability. The current whole-genome approach, which has been validated in the human genome sequencing project, uses BAC end sequences and restriction fingerprints to build BAC contigs. Restriction fingerprints not only serve as independent confirmation of BAC contigs assembled based on BAC end sequence, but also enable identification of chimeric BACs and clones with internal deletions. This map-as-you-go strategy saves substantial time and effort in constructing sequence ready maps. This proposal is to perform BAC end sequencing at TIGR and restriction fingerprinting at the Clemson University Genomics Institute Physical Mapping Center from the mouse BAC library or libraries constructed to support mouse genome sequencing. The clones, the sequences, gel images of the fingerprints, and the binning of the clones using the FPC program (C. Soderlund, Sanger Centre) will be an available resource for those sequencing the mouse genome, and the community at large. We will continue our focus on quality in this BAC clone end sequencing and restriction fingerprinting project while developing and implementing automation and new methodologies for reducing costs and increasing throughput.

随着人类项目转移到生产测序中，一个计划正在制定测序小鼠基因组。 当前的计划采用“首先，映射第二”策略，与人类项目一样，基于BAC克隆的shot弹枪测序，其中包括所选小鼠菌株基因组（C57BL6/J）的基因组的连续区域。在细菌人造染色体（BAC）载体中构建的库已成为高通量基因组测序项目的选择，因为它们的克隆能力和高稳定性。 当前在人类基因组测序项目中验证的当前全基因组方法使用BAC最终序列和限制指纹来构建BAC重叠群。限制指纹不仅是基于BAC末端序列组装的BAC重叠群的独立确认，而且还可以鉴定具有内部缺失的嵌合BAC和克隆。 此地图策略可以节省大量时间和精力来构建序列现成的地图。该建议是在TIGR上进行BAC结束测序，并在Clemson University基因组学研究所的物理映射中心或构建的库中限制指纹识别，或者构建了用于支持小鼠基因组测序的库。 使用FPC程序（C. Soderlund，Sanger Center）的克隆，指纹的序列，凝胶图像以及克隆的框架将是对那些测序鼠标基因组和整个社区的人的可用资源。 在开发和实施自动化以及降低成本和增加吞吐量的新方法的同时，我们将继续关注BAC克隆端测序和限制指纹项目的质量。