| 莱芜猪心脏cDNA文库的构建与鉴定 |
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| 引用本文： | 李付美,闫顺,曲志才,徐来祥. 莱芜猪心脏cDNA文库的构建与鉴定[J]. 生物技术通报, 2010, 0(3) |
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| 作者姓名： | 李付美 闫顺 曲志才 徐来祥 |
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| 作者单位： | 曲阜师范大学生命科学学院,曲阜,273165 |
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| 基金项目： | 国家科技基础条件平台建设项目 |
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| 摘 要： | 以莱芜猪心脏为供试材料,采用改进的异硫氰酸胍法提取心脏总RNA。通过SMART技术反转录合成全长cD-NA,经LD-PCR扩增后与pMD18-T载体连接并转化细菌DH5α,测定滴度和重组率,构建成莱芜猪心脏全长cDNA文库。经初步检测,原始文库的滴度为1.8×105CFU/mL,重组率约为94.4%。从原始文库随机挑取13个单菌落过夜培养,提取质粒,经HindⅢ和EcoRⅠ双酶切,凝胶电泳显示插入片段大小在0.3-2.0kb之间。
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| 关 键 词： | 莱芜猪 心脏 cDNA文库 SMART技术 |
Construction and Identification of cDNA Library of the Heart tissue in Laiwu Pig |
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| Li Fumei,Yan Shun,Qu Zhicai,Xu Laixiang. Construction and Identification of cDNA Library of the Heart tissue in Laiwu Pig[J]. Biotechnology Bulletin, 2010, 0(3) |
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| Authors: | Li Fumei Yan Shun Qu Zhicai Xu Laixiang |
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| Affiliation: | Li Fumei Yan Shun Qu Zhicai Xu Laixiang(College of Life Science,Qufu Normal University,Qufu 273165) |
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| Abstract: | The total RNA from heart tissue of Laiwu pig was extracted by isothiocyanate method Full-length cDNAs were synthesized by SMART technique and a pool of double-stranded cDNA was amplified by LD-PCR assay The recovered cDNA fragments were ligated into pMD18-T vector and transformed E coli DH5α to construct a full-length cDNA library The library was tittered,and the rate of recombination(blue/white)was determined The results showed that the primary titer of the constructed cDNA library was 1.8×10~5 CUF/mL,the recombination rate was about 94 4% and the length of most cDNA inserted in the library was about 0.3-2.0 kb by agarose gel electrophoresis. |
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| Keywords: | Laiwu pig Heart tissue cDNA library SMART technique |
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