SEMI-MULTIPLEX PCR TECHNIQUE FOR SCREENING OF ABUNDANT TRANSCRIPTS DURING SYSTEMATIC SEQUENCING OF CDNA LIBRARIES

The systematic sequencing of cDNA libraries is an efficient approach f or the identification of new genes, but the presence of abundant mRNAs is often a major problem. This paper describes a very simple method o f ''semi-multiplex PCR'' that allows specific identification of such a bundant transcripts before DNA sequencing without using nonrepresentat ive subtracted libraries. The PCR utilizes a series of forward primers specific for abundant transcripts with a pair of universal primers us ed for template generation. cDNA clones corresponding to abundant mRNA s are then revealed by double bands in agarose gel.