Semi-Multiplex PCR Technique for Screening of Abundant Transcripts During Systematic Sequencing of cDNA Libraries. (October 1996)

• Record Type: Journal Article
• Authors: Pacchioni, B.; Trevisan, S.; Gomirato, S.; Toppo, S.; Valle, G.; Lanfranchi, G.
• Is Part Of: Biotechniques. Volume 21:Number 4(1996)
• Page Start: 644
• Page End: 649
• Publication Date: 1996-10
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/96214st01
• Languages: English
• ISSNs: 0736-6205

Abstract

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.