cDNA+末端快速扩增技术(+RACE)+的优化与改良.pdfVIP

7 3 生命科学研究 Vol. 7 No. 3 2003 9 Life Science Rese rch Sep. 2003 cDNA ( RACE) 1, 2 2 2 2 李关荣 , 鲁 成, 夏庆友 , 向仲怀 ( 1. , 400716; 2. , 400716) : cDNA 末端的快速扩增( RACE) 法是延伸已知部分外显子序列 克隆全长 cDNA 基因的主要方法之 一. 广泛用于许多已知功能基因片段的进一步延伸 全长cDNA 的克隆. 但由于其过程复杂涉及多步连续的 酶促过程, 如反转录TdT 加尾第二链合成RACE- PCR 扩增及RACE 产物克隆等, 在实际应用中有许多问题 相当难度. 主要针对RACE 各步骤中存在的问题进行了分析, 并对其优化 改良进行了综述. : cDNA 末端的快速扩增法; 优化; 改良 : Q781 : A : 1007-7847( 2003) 03-0189-09 The Optimization and Improvement of Rapid Amplification of cDNA Ends ( RACE) 1, 2 2 2 2 LI Gu n-rong , LU Cheng , XIA Qing-you , XIANG Zhong-hu i ( 1. College of Agronomy and Lfi e Sciences, Southwest Agricultural University, Chongqing 400716, China; 2. College of Sericulture and Biotechnology, Southwest Agricultural University, Chongqing 400716, China) Abstract: R pid mplific tion of cDNA ends ( RACE) is one of the m in methods for extending p rti lly known exon sequence nd cloning of its ful-l length cDNA. It h s been widely used in further extending of function l fr gments of genes of interests especi lly the cloning of the full-length cDNAs. RACE involves multiple enzyme- c t lyzed processes, such s reverse tr nscription, TdT t iling, the second-str nd cDNA synthesis, RACE-PCR mplific tion nd the cloning of RACE products, which in pr ctice h ve m ny problems nd difficulties. These problems re n lyzed in depth nd the optimiz tions nd improvements on e ch of the processes re reviewed. Key words: RACE; optimiz tion; improvement