cDNA末端快速扩增技术的研究进展

cDNA末端快速扩增技术是一种基于多聚酶链式反应的技术 ,它的发展大大便利了应用其它方法获得的部分cDNA序列后克隆全长cDNA 5’和 3’末端的工作。不仅RACE方法能在短时间内得到完整的cDNA末端序列 ,而且一些截短的cDNA末端常常也能在RACE的过程中被扩增 ,而这些截短的产物破坏了全长cDNA克隆的获取。许多研究者对RACE的流程提出了改进方案 ,从而提高了该技术的效力。本文介绍了许多已发表的RACE技术关键步骤的改良 ,包括一些具体有效的操作流程 ,如RNA连接酶介导的RACE/连接锚定PCR等 ,还有其有效性的例证。

Development of Rapid Amplification of cDNA Ends

Rapid amplification of cDNA ends (RACE) is a polymerase chain reaction (PCR)-based technique which was developed to facilitate the cloning of full-length cDNA 5' and 3' ends after a partial cDNA sequence has been obrained by other methods. While RACE can yield complete sequences of cDNA ends in only a few days, the procedure frequently results in the exclusive amplification of truncated cDNA ends, undermining efforts to generate full-length clones. Many investigators have suggested modifications to the RACE protocol to improve the effectiveness of the technique. The review presents numerous published variations of the key steps in the RACE method. Also included is a detailed, effective protocol of RNA ligase-mediated RACE/ligation-anchored PCR.