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Construction and targeted retrieval of specific clone from a non-gridded soybean bacterial artificial chromosome library
Authors:Zhengjun Xia  Hongyan Wu  Satoshi Watanabe  Kyuya Harada
Affiliation:1. Key Laboratory of Soybean Molecular Design Breeding, Northeast Institute of Geography and Agroecology, Chinese Academy of Sciences, Harbin 150081, China;2. Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;3. National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan
Abstract:Although a post-genomic era is emerging for many plants, the bacterial artificial chromosome (BAC) library is still a valuable tool for genomic studies and preservation of precious genetic resources. Construction of non-gridded BAC libraries would dramatically reduce cost and save storage space. A non-gridded BAC library composed of approximately 96,000 insert-containing clones in 80 pools with an average insert size of 75 kb was constructed. This library represented 5.2 genome equivalents. We successfully developed a unique procedure to retrieve positive clones from the non-gridded pools. With this retrieving protocol, the non-gridded library system can be adapted to different species and to serve various research needs.
Keywords:BAC   Pooling   Screening   Non-gridded library   Soybean
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