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A simple and reliable 5'-RACE approach

2000; Oxford University Press; Volume: 28; Issue: 22 Linguagem: Inglês

10.1093/nar/28.22.e96

1362-4962

G. Schramm,

Genomics and Phylogenetic Studies

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.