A rapid DNA assembling strategy mediated by direct full-length polymerase chain reaction |
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| Authors: | Lu-Bin Zhou Qing-Qing Lin Jing-Xian Zhang Shu-Juan Zhao Zhi-Bi Hu |
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| Affiliation: | 1. The SATCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, the MOE Key Laboratory for Standardization of Chinese Medicines and Shanghai Key Laboratory of Complex Prescription, Institute of Chinese Material Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;2. The Key Laboratory of Modern Chinese Medicines, School of Chinese Pharmacy, China Pharmaceutical University, Nanjing 210009, China |
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| Abstract: | An efficient DNA assembling strategy was developed here modified from Class-IIS endonuclease mediated DNA splicing by directed ligation (SDL). Benefited from the full-length PCR directly using ligation products as template, this strategy required less effort and less time to obtain the assembled full-length DNA. The advantages of this strategy made it a rapid and easy-to-perform gene splicing and multiple site-directed mutagenesis approach especially practicable when more fragments need to be assembled at the same time. |
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| Keywords: | AnSh, Aspergillus niger salicylate hydroxylase OE-PCR, over extension PCR PCR, polymerase chain reaction SDL, DNA splicing by directed ligation SDM, site-directed mutagenesis |
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