下载后可任意编辑3 种 5’RACE 技术的比较与优化 【摘要】 目的 讨论环化法、末端脱氧核糖核酸转移酶法和 SMART 反转录酶法 3种常用 5‘RACE 的技术的特点，并对它们进行了优化。方法 以播娘蒿 RNA 为模板，先按文献标准方法进行 5‘RACE，然后通过增加反应时间，提高部分反应物浓度等方法进行优化。结果 未优化前环化法和 TdT酶法条带几乎不可见，SMART 反转录酶法隐下载后可任意编辑约可见条带，但不清楚。优化条件后，环化法出现弥散条带，TdT 酶法胜之，但条带依旧弥散，而 SMART 反转录酶法最佳，获得清楚特异性条带。结论 SMART 5‘RACE效果最好，其次为 TdT 酶法，环化法效果有待改进，同时通过优化条件可以明显增加产物的特异性，为实验讨论中 5‘RACE方法的选择提供了依据。【关键词】 5‘RACE；比较；优化 Abstract ： Objective To study the characters of Cycling 5‘RACE,TdT 下载后可任意编辑5‘RACE and SMART 5‘RACE methods and optimize Standard 5‘RACE ways were done at first with RNA templet of Descurainia they were optimized with delaying the reaction time and increasing the concentrations of some Before optimized,the gel-straps in Cycling 5‘RACE and TdT 5‘RACE were gel-strap in SMART 5‘RACE could be found but not optimized,dispersed gel-straps appeared in Cycling 下载后可任意编辑5‘RACE,the result of TdT 5‘RACE was better and still best was SMART 5‘RACE,which got the clear and specific The result of SMART 5‘RACE was best,TdT 5‘RACE was better,and Cycling 5‘RACE way was still under optimization,the specificity of the results could be obviously provided bases for choosing of 5‘RACE methods in research. Key 下载后可任意编辑words:5‘RACE;comparison;eptimization 经典克隆未知基因的方法比如通过筛选文库等有个共同的弊病，即实验操作繁琐，周期较长、工作量繁重，且不易得到全长cDNA 序列。近年来随着 PCR 技术的快速兴起和成熟，cDNA 末端快速扩增技术(Rapid Amplification of cDNA Ends,RACE) 为 简洁方便的基因克隆进展道路指出了新的方向。RACE 是利用 PCR 技术由已知的...
