CONSTRUCTION OF CDNA LIBRARIES FROM SMALL AMOUNTS OF TOTAL RNA USING THE SUPPRESSION PCR EFFECT

Here we describe a method for preparing high-quality cDNA libraries fr om total RNA. By this method, double-stranded (ds) cDNA ligated with a specially designed ds adaptor is amplified by PCR using a modified T- primer and another primer corresponding to the outer part of the adapt or, The suppression PCR effect strongly inhibits the amplification of poly(A)- RNA, thereby reducing background, This method leads to amplif ication of high-quality cDNA, facilitating the construction of represe ntative cDNA libraries from as little as 10-100 ng of total RNA. (C) 1 997 Academic Press