Luo Meizhong, Wing Rod A
Department of Plant Sciences, Arizona Genomics Institute, University of Arizona, Tucson, AZ, USA.
Methods Mol Biol. 2003;236:3-20. doi: 10.1385/1-59259-413-1:3.
Large genomic DNA insert-containing libraries are required as critical tools for physical mapping, positional cloning, and genome sequencing of complex genomes. The bacterial artificial chromosome (BAC) cloning system has become a dominant system over others to clone large genomic DNA inserts. As the costs of positional cloning, physical mapping, and genome sequencing continuously decrease, there is an increasing demand for high-quality deep-coverage large insert BAC libraries. In our laboratory, we have constructed many high-quality deep-coverage large insert BAC libraries including arabidopsis, manocot and dicot crop plants, and plant pathogens. Here, we present the protocol used in our laboratory to construct BAC libraries.
含有大片段基因组DNA插入片段的文库是复杂基因组物理图谱构建、定位克隆和基因组测序的关键工具。细菌人工染色体(BAC)克隆系统已成为克隆大片段基因组DNA插入片段的主导系统。随着定位克隆、物理图谱构建和基因组测序成本的不断降低,对高质量、深度覆盖的大片段插入BAC文库的需求日益增加。在我们实验室,我们已经构建了许多高质量、深度覆盖的大片段插入BAC文库,包括拟南芥、单子叶和双子叶作物以及植物病原体。在此,我们介绍我们实验室用于构建BAC文库的方案。