循环逆转录反应方法及应用条件研究  

Repeated Reverse Transcription Reaction (RRTR) Protocol and Its Application

作　　者：陈燃[1] 金詟[1] 伍迪[1] 唐榕[1] 毛裕民[1] 

机构地区：[1]复旦大学生命科学学院遗传所遗传工程国家重点实验室,上海200433

出　　处：《生物化学与生物物理进展》2001年第4期560-562,共3页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金资助项目 (3 970 0 0 82 )~~

摘　　要：建立了循环逆转录反应方法 (repeatedreversetranscriptionreaction ,RRTR) .其原理是通过控制高温变性 低温退火延伸的循环过程 ,反复利用起始RNA模板进行逆转录反应 .RNA杂交的结果证明在RRTR过程中 ,RNA模板是保持稳定的 ;点杂交结果表明cDNA产物量得到了增加 ;定量PCR的结果表明RRTR是cDNA线性增长过程 .结果提示 ,RRTR可有效地提高RNA检测灵敏度 。The RRTR can linearly increase cDNA products via repeated reverse transcription reactions controlled by temperatures and catalyzed by FD TRT (FD thermostable reverse transcriptase). The stability of RNA in RRTR is proved via the Northern blot, and the increase in cDNA products along with that in reacting cycles is shown via the Dot hybridization. After components of the reaction system are optimized, the RRTR combined with PCR is applied in quantification of HCV RNA through changing the reacting cycles of the RRTR. It can be concluded that due to the linearity of RRTR, the RRTR is a convenient method to enhance the sensitivity and efficacy of RNA related study. It can be used in some fields which full length cDNA is not required, such as quantification of RNA or cDNA, RACE (rapid amplification of cDNA end), and different display.

关 键 词：循环逆转录反应 RNA 定量分析 应用 

分 类 号：Q522[生物学—生物化学]