A simple and reliable 5'-RACE approach

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA, Complementary / genetics*
  • Entamoeba / genetics
  • Entamoeba histolytica / genetics
  • Polymerase Chain Reaction / methods*
  • RNA, Protozoan / genetics

Substances

  • DNA, Complementary
  • RNA, Protozoan