梅花鹿心脏组织cDNA文库的构建

Construction of cDNA Library from the Heart of Sika Deer

[目的]构建梅花鹿心脏cDNA文库。[方法]提取梅花鹿心脏总RNA,纯化mRNA;合成双链cDNA,连接载体,并电转化至感受态细胞;测定文库的克隆数、重组率及插入片段大小。[结果]经鉴定,所构建梅花鹿心脏组织cDNA文库,库容量为3.8×106个克隆,重组率为100%,插入片段大小为0.5～2.0kb,平均长度约为1.0kb。[结论]所构建cDNA文库的各项指标均达到要求。

[Objective]To construct a cDNA library from the heart of sika deer.[Method]Total RNA was extracted from the heart of sika deer,and mRNA was purified.Double-stranded cDNA was synthesized,then connected with vector and electrotransformated to competent cells.The number of clones,recombinant rate and the length of insert fragments were checked.[Result]The capacity of the cDNA library was 3.8×106 with a recombinant rate of 100%.The length of insert fragments ranged from 0.5 kb to 2.0 kb,and the average length was about 1.0 kb.[Conclusion] The cDNA library constructed was qualified.