Molecular Biotechnology

, Volume 27, Issue 3, pp 179–186 | Cite as

RACE using only a gene-specific primer

Application of a template-switching model
Research

Abstract

This article describes a simple method for accurate rapid amplification of complementary deoxyribonucleic acid (cDNA) ends (RACE), the distinctive feature being that only a gene-specific primer is used, without an anchor or adapter primer. Under these conditions, Thermus aquaticus (Taq) polymerase synthesizes cDNA ends exactly, so that amplified products obtain a characteristic structure: a terminal inverted repeat composed of a gene-specific primer and occasionally several nucleotides from its 3′ flanking sequence. These structures suggest a hypothetical mechanism of cDNA end synthesis in which Taq DNA polymerase synthesizes a sequence complementary to the gene-specific primer at the 3′ end of the daughter strand by switching the template to the 5′ terminal region through circularization of the DNA. As a result, the targeted cDNA will be efficiently amplified with only a single gene-specific primer. This technique, which provides highly specific amplification of the 5′ and 3′ ends of a cDNA, is especially useful for isolation of cDNA when the corresponding messenger ribonucleic acid is scarce.

Index Entries

RACE single gene-specific primer terminal inverted repeat template-switching model of Taq DNA polymerase 

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Copyright information

© Humana Press Inc. 2004

Authors and Affiliations

  1. 1.HuBit GenomixTokyoJapan

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