Department of Soil and Crop Sciences, Texas A&M University, College Station, Texas, USA.
Nat Protoc. 2012 Feb 16;7(3):479-99. doi: 10.1038/nprot.2011.456.
Large-insert BAC (bacterial artificial chromosome) and BIBAC (binary BAC) libraries are essential for modern genomics research for all organisms. We helped pioneer the BAC and BIBAC technologies, and by using them we have constructed hundreds of BAC and BIBAC libraries for different species of plants, animals, marine animals, insects, algae and microbes. These libraries have been used globally for different aspects of genomics research. Here we describe the procedure with the latest improvements that we have made and used for construction of BIBAC libraries. The procedure includes the preparation of BIBAC vectors, the preparation of clonable fragments of the desired size from the source DNA, the construction and transformation of BIBACs and, finally, the characterization and assembly of BIBAC libraries. We also specify the modifications necessary for construction of BAC libraries using the protocol. The entire protocol takes ∼7 d.
大片段细菌人工染色体 (BAC) 和双元 BAC (BIBAC) 文库是所有生物体现代基因组学研究的重要基础。我们率先开发了 BAC 和 BIBAC 技术，并利用这些技术为不同植物、动物、海洋动物、昆虫、藻类和微生物物种构建了数百个 BAC 和 BIBAC 文库。这些文库已被全球用于基因组学研究的各个方面。本文描述了我们进行 BIBAC 文库构建时所使用的最新改进方法。该方法包括 BIBAC 载体的准备、从源 DNA 中制备所需大小的可克隆片段、BIBAC 的构建和转化，以及最终 BIBAC 文库的特征和组装。我们还指定了使用该方案构建 BAC 文库所需的修改。整个方案大约需要 7 天。
