Semi-Multiplex PCR Technique for Screening of Abundant Transcripts During Systematic Sequencing of cDNA Libraries

oleh: B. Pacchioni, S. Trevisan, S. Gomirato, S. Toppo, G. Valle, G. Lanfranchi

Format: Article
Diterbitkan: Future Science Ltd 1996-10-01

Deskripsi

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