BAC Library Production for Comparative Genetics

目标是为许多物种准备高质量的BAC文库，以满足科学界对比较基因组学和功能应用的兴趣。这项工作将由两个合作小组完成，一个由劳伦斯伯克利国家实验室（LBNL）的郑博士领导，另一个由奥克兰儿童医院研究所的陈博士共同领导。Osoegawa & deJong。该应用程序包括与制备细菌人工染色体（BAC）相关的四个部分：1)BAC文库制备，2)克隆阵列和文库复制，3)文库表征，以及4)研究与开发，以提高整体流程效率和BAC文库质量。在第一年，该联盟将生成并表征12个具有10倍基因组冗余的动物BAC文库。假定对更多gbac的需求增加和生产效率的提高，将分别在第2年和第3年编制和鉴定17和22个额外的文库。这些文库基于目前的BAC克隆技术，将补充和扩展目前可用于比较基因组分析的少量BAC克隆库。新的克隆收集将通过一组标准化测试进行分析，包括使用一组经济标记进行筛选、有限BAC末端测序、BAC指纹图谱、BAC稳定性分析和插入物大小确定。特别重要的是质量测试，以确保低BAC稳定性分析和插入尺寸确定。特别重要的是质量测试，以确保低水平的克隆交叉污染。一旦库通过了质量控制，新的资源将以与主要应用程序一致的格式提供。为了提高整个过程的效率，将制定一套标准操作程序（SOP），使BAC克隆从一种艺术形式转向常规的高通量程序。预计新的BAC资源将有助于常规高通量程序。期望新的BAC资源将有助于通过比较动物物种光谱中的基因来更好地了解基因功能和进化。BAC克隆也将成为创建人类疾病动物模型的工具。广泛传播克隆是这项建议的一个没有经费但必不可少的组成部分。图书馆信息将通过出版物和我们的主页（http://www.chori.org/bacpac）传播。

The goal is to prepare BAC libraries for many species, of high quality, tailored to the scientific community's interest in comparative genomics and functional applications. The work will be done by two collaborating teams, one lead by Dr. Cheng at the Lawrence Berkeley National Laboratory (LBNL) and the other one at the Children's Hospital Oakland Research Institute, co-directed by Drs. Osoegawa & deJong. The application has four components relevant to preparing Bacterial Artificial Chromosomes (BACs): 1) BAC Library preparation, 2) Clone Arraying & Library Duplication, 3) Library Characterization, and 4) Research & Development to improve the overall process efficiency and the BAC library quality. During the first year, the consortium will generate and characterize twelve animal BAC libraries with ten-fold genome redundancy. Presuming an increasing need for additional GBACs and improved production efficiency, 17 and 22 additional libraries will be prepared and characterized in years 2 and 3 respectively. The libraries, based on current BAC cloning technology, will complement and extend the small repertoire of BAC clone collections now available for comparative genome analysis. The new clone collections will be analyzed by a standardized set of tests, including screening with a set of economic markers, limited BAC-end sequencing, BAC fingerprinting, BAC stability analysis and insert size determination. Of particular importance are quality tests to ensure low BAC stability analysis and insert size determination. Of particular importance are quality tests to ensure low levels of clonal cross contamination. Once the libraries pass the quality controls, the new resources will be made available in a format consistent with major applications. To improve the overall process efficiency, a set of Standard Operating Procedures (SOP's) will be developed to move BAC cloning away from an art form towards a routine high-throughput procedures. It is expected that the new BAC resources will contributed towards a routine-high throughput procedure. It is expected that the new BAC resources will contribute to better understanding of gene function and evolution through comparison of genes in a spectrum of animal species. The BAC clones will also become tools to create animal models for human diseases. Widespread dissemination of the clones is an unfunded but essential component of this proposal. Library information will be disseminated by publication and through our home page (http://www.chori.org/bacpac).