A simple and reliable 5'-RACE approach
Open Access
- 15 November 2000
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 28 (22) , 96e-96
- https://doi.org/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.Keywords
This publication has 7 references indexed in Scilit:
- CapSelect: a highly sensitive method for 5' CAP-dependent enrichment of full-length cDNA in PCR-mediated analysis of mRNAsNucleic Acids Research, 1999
- Amplification of cDNA ends based on template-switching effect and step- out PCRNucleic Acids Research, 1999
- Solid-phase cDNA library construction, a versatile approach.Nucleic Acids Research, 1998
- Revolutions in Rapid Amplification of cDNA Ends: New Strategies for Polymerase Chain Reaction Cloning of Full-Length cDNA EndsAnalytical Biochemistry, 1995
- PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates.Proceedings of the National Academy of Sciences, 1994
- A highly sensitive method for mapping the 5′ termini of mRNAsNucleic Acids Research, 1993
- Differential Display of Eukaryotic Messenger RNA by Means of the Polymerase Chain ReactionScience, 1992