Optical mapping of a rice BAC clone using restriction endonuclease and imaging with fluorescent microscopy at single molecule level |
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Authors: | Zhenqian Ouyang Shiquan Wang Wenxue Zhai Lihuang Zhu |
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Affiliation: | (1) Institute of Genetics and Development of Chinese Academy of Sciences, 100101 Beijing, China;(2) Rice Institute at Sichuan Agriculture University, 611130 Chengdu, China |
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Abstract: | A method of constructing restriction map by optical mapping and single molecule fluorescent microscopy is described. DNA molecules were aligned and adsorbed on a glass coverslip surface by a modified “molecular combing” technique, and then the surface-immobilized DNAs were cleaved in situ with a restriction endonuclease. Individual DNA molecules digested by the endonuclease EcoR I were observable with fluorescent microscopy. Using optical mapping, a physical map of a rice bacterial artificial chromosome clone was constructed. This method will facilitate genomic mapping and tracing the dynamic process in real time at a single molecule level with fluorescence microscopy. |
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Keywords: | digesting DNA in situ single molecule detecting optical mapping physical map BAC |
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