Juntang Lin, Pramanik Jogenananda, Congrui Wang, Huiyong Zhang, Huigen Feng, Baosheng Yang, Yuchang Li, Cunshuan Xu
Department of Cell Biology, Xinxiang Medical College, 453003 Xinxiang, China.
Indian J Clin Biochem. 2004 Jul;19(2):181-3. doi: 10.1007/BF02894282.
The study aims to construct cDNA library of Changliver cell by SMART (switching mechanism at 5' end of RNA transcript) technique and analyze its quality. cDNA of Changliver cell was made with RT-PCR and LD-PCR (long-distance PCR), the cDNA library was constructed with SMART cDNA library construction kit. Through testing, the high quality cDNA library containing whole long cDNA of Changliver cell had been constructed. The titer of the amplified cDNA library was 4.5 × 10(10) pfu/ml and the average exogenous inserts of the recombinants is 1.5 kb. These results showed that the Changliver cell cDNA library had an excellent quality and lay foundation for screening whole long cDNA of related genes.
本研究旨在通过SMART（RNA转录本5'端切换机制）技术构建Changliver细胞的cDNA文库并分析其质量。采用RT-PCR和LD-PCR（长距离PCR）制备Changliver细胞的cDNA，使用SMART cDNA文库构建试剂盒构建cDNA文库。经检测，成功构建了包含Changliver细胞全长cDNA的高质量cDNA文库。扩增后的cDNA文库滴度为4.5×10(10)pfu/ml，重组子的平均外源插入片段为1.5kb。这些结果表明Changliver细胞cDNA文库质量优良，为筛选相关基因的全长cDNA奠定了基础。
