A simple and reliable 5'-RACE approach
A simple and reliable 5'-RACE approach
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DOI：
10.1093/nar/28.22.e96
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发表时间：
2000-11-15
影响因子：
14.9
通讯作者：
Roeder, T
中科院分区：
文献类型：
--
作者：
Schramm, G;Bruchhaus, I;Roeder, T
描述了一种新的扩增cDNA端的方法。它只需要极少量的材料，一个简单的cDNA合成反应和一个单一的PCR反应就可以扩增出特定目的基因的5‘或3’端。它将所谓的CapFinder方法与固相cDNA合成相结合，从而几乎消除了通常与5‘-RACE协议相关的背景问题。该方法只需一次合成反应即可获得多个cDNA的完整5‘端。与LA聚合酶链式反应相结合，可扩增出几千个碱基的未知5‘末端。它易于执行，快速，廉价和可靠，这应该使它能够取代目前使用的大多数5‘-RACE协议
A novel approach for the amplification of cDNA ends is described. It requires only minimal amounts of material, a simple cDNA synthesis reaction and a single PCR reaction to amplify the desired 5'- or 3'-ends of a certain cDNA of interest. It combines the so called CapFinder approach with solid phase cDNA synthesis, thus almost eliminating background problems usually associated with 5'-RACE protocols. This approach could be used to generate complete 5'-ends of numerous cDNAs using only one cDNA synthesis reaction. In combination with LA PCR, several kilobases of unknown 5'-ends could be amplified. It is easy to perform, quick, inexpensive and reliable, which should enable it to replace most currently used 5'-RACE protocols