当前位置: 首页 > 文章 > 莱芜猪心脏cDNA文库的构建与鉴定 生物技术通报 2010,0 (3) 139-142
Position: Home > Articles > Construction and Identification of cDNA Library of the Heart tissue in Laiwu Pig Biotechnology Bulletin 2010,0 (3) 139-142

莱芜猪心脏cDNA文库的构建与鉴定

作  者:
李付美;闫顺;曲志才;徐来祥
单  位:
曲阜师范大学生命科学学院
关键词:
莱芜猪;心脏;cDNA文库;SMART技术
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。
译  名:
Construction and Identification of cDNA Library of the Heart tissue in Laiwu Pig
作  者:
Li Fumei Yan Shun Qu Zhicai Xu Laixiang(College of Life Science,Qufu Normal University,Qufu 273165)
关键词:
Laiwu pig Heart tissue cDNA library SMART technique
摘  要:
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 1.8×105 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.

相似文章

计量
文章访问数: 13
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊