OPTIMIZATION PROCEDURE FOR OLIGONUCLEOTIDES AND AMPLIFICATION CONDITIONS BY PCR FOR RACE (5 'AND 3'-RAPID AMPLIFICATION OF CDNA ENDS).

摘要

Optimization procedure for oligonucleotides and amplification conditions by PCR for RACE (5 '' and 3 '' - rapid amplification of cDNA ends). The present invention describes a method for optimizing oligonucleotides and PCR amplification conditions used for RACE, by cloning a gene of interest / problem cDNA fragment into a genetic construct that includes the sequences corresponding to the 5''-RACE oligonucleotides and 3 '' - RACE flanking the cloning site of the gene of interest / problem cDNA fragment. This procedure makes it possible to check the compatibility of the oligonucleotides necessary for the amplification of the gene of interest / fragment of the problem cDNA with the oligonucleotides 5 '' - RACE and 3 '' - RACE and the optimal conditions of the PCR reaction; acting in this way as a control process prior to the RACE experiment itself.