A simple and reliable 5'-RACE approach.
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.
Schramm
G
G
Universität Hamburg Zoologisches Institut, Martin-Luther-King-Platz 3, D-20146 Hamburg, Germany.
Bruchhaus
I
I
Roeder
T
T
D000818:Animals; D018076:DNA, Complementary / Q000235:genetics*; D004747:Entamoeba / Q000235:genetics; D004748:Entamoeba histolytica / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D016053:RNA, Protozoan / Q000235:genetics
D018076:DNA, Complementary; D016053:RNA, Protozoan