A QuikChange-like method to realize efficient blunt-ended DNA directional cloning and site-directed mutagenesis simultaneously
文献类型：期刊论文
| 作者 | An, Yingfeng; Lv, Anguo; Wu, Wenfang |
| 刊名 | BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
 |
| 出版日期 | 2010-06-25 |
| 卷号 | 397期号:2页码:136-139 |
| 关键词 | Quikchange Kit Site-directed Mutagenesis Blunt-ended Dna Ligation Mutant Library |
| ISSN号 | 0006-291X |
| DOI | 10.1016/j.bbrc.2010.05.042 |
| 英文摘要 | Here we present a QuikChange-like method to efficiently realize blunt-ended DNA cloning and conveniently introduce a site-directed mutation to recombinant plasmid at the same time. After blunt-ended DNA ligation and transformation, the plasmid DNA mixture is extracted from pooled transformants and directly used as template for PCR amplification with a pair of complementary mutagenic primers. With this method, sam1 gene was inserted into pUC19 vector by blunt-end ligation, and a unique restriction site Spe 1 was introduced to the recombinant plasmid at the same time. The randomly selected transformants were analyzed by DNA sequencing, and most of the clones were found to have correct sequences. However, no correct construct was found from randomly selected transformants after traditional blunt-ended DNA ligation and transformation. (C) 2010 Elsevier Inc. All rights reserved. |
| WOS研究方向 | Biochemistry & Molecular Biology ; Biophysics |
| 语种 | 英语 |
| WOS记录号 | WOS:000279718900003 |
| 出版者 | ACADEMIC PRESS INC ELSEVIER SCIENCE |
| 源URL | [http://210.72.129.5/handle/321005/121397]  |
| 专题 | 中国科学院沈阳应用生态研究所 |
| 通讯作者 | Wu, Wenfang |
| 作者单位 | Chinese Acad Sci, Inst Appl Ecol, Shenyang 110016, Peoples R China |
| 推荐引用方式 GB/T 7714 | An, Yingfeng,Lv, Anguo,Wu, Wenfang. A QuikChange-like method to realize efficient blunt-ended DNA directional cloning and site-directed mutagenesis simultaneously[J]. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,2010,397(2):136-139. |
| APA | An, Yingfeng,Lv, Anguo,&Wu, Wenfang.(2010).A QuikChange-like method to realize efficient blunt-ended DNA directional cloning and site-directed mutagenesis simultaneously.BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS,397(2),136-139. |
| MLA | An, Yingfeng,et al."A QuikChange-like method to realize efficient blunt-ended DNA directional cloning and site-directed mutagenesis simultaneously".BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 397.2(2010):136-139. |
入库方式： OAI收割
来源：沈阳应用生态研究所
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