摘要: |
用RNA提取试剂——TRIZOL Reagent提取刺参(Apostichopus japonicus)肌肉组织总RNA,用SMART cDNA Library Construction Kit构建cDNA文库。经测定原始文库滴度达到 3.2×106,扩增后文库滴度达到 5.1×109,重组率达到96.7%,从扩增文库随机挑取12 个克隆进行PCR 扩增鉴定,结果显示,插入片段大小为0.5~2.5 kb。通过各项指标验证成功构建了刺参肌肉组织cDNA 文库。
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关键词: 刺参(Apostichopus japonicus) 肌肉组织 cDNA文库 |
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基金项目:国家自然科学基金项目(30571417) |
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Construction of cDNA library from the musculature of Apostichopus japonicus |
CHEN Lu,JIANG Guo-liang,LIU Yun,QIU Lei,XIANG Peng
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Abstract: |
Total RNA of muscles of Apostichopus japonicus were extracted by TRIZOL Reagent. cDNA library was constructed with SMART cDNA library construction kit. The primary library hada high titer of 3.2×106, in which 96.7% clones were recombinant. The amplified library hada titer of 5.1×109, and the length of most cDNA inserted in the library was about 0.5~2.5 kb. It was concluded that the cDNA library of Musculature of A. japonicus had been constructed by using the SMART technique.
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Key words: Apostichopus japonicus muscle cDNA library |