一种简单可靠的5'-RACE方法。
A simple and reliable 5'-RACE approach.
作者信息
Schramm G, Bruchhaus I, Roeder T
机构信息
Universität Hamburg Zoologisches Institut, Martin-Luther-King-Platz 3, D-20146 Hamburg, Germany.
出版信息
Nucleic Acids Res. 2000 Nov 15;28(22):E96. doi: 10.1093/nar/28.22.e96.
Abstract
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.
摘要
本文描述了一种用于扩增cDNA末端的新方法。它仅需要极少量的材料、一个简单的cDNA合成反应和一个单一的PCR反应，即可扩增特定感兴趣cDNA的所需5'或3'末端。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方案相关的背景问题。该方法可仅使用一个cDNA合成反应来生成众多cDNA的完整5'末端。与LA PCR相结合，可以扩增几千个碱基的未知5'末端。它易于操作、快速、廉价且可靠，这应该使其能够取代目前大多数使用的5'-RACE方案。
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本文引用的文献
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