Generation of High-Quantity and Quality Tag/Ditag cDNAs for SAGE Analysis. (August 2001)

• Record Type: Journal Article
• Authors: Lee, Sanggyu; Chen, Jianjun; Zhou, Guolin; Wang, San Ming
• Is Part Of: Biotechniques. Volume 31:Number 2(2001)
• Page Start: 348
• Page End: 354
• Publication Date: 2001-08
• 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: 10.2144/01312st07
• Languages: English
• ISSNs: 0736-6205

Abstract

The serial analysis of gene expression (SAGE) technique is an important tool for genome-wide gene expression analysis. However, the requirement of a large amount of mRNA for the analysis and the difficulties in generating high-quality tag and ditag fragments for the construction of a SAGE library often interfere with the successful performance of the SAGE technique. We developed two procedures to solve these issues: (i) introducing low-cycle PCR amplification of the 3′ cDNA before the Bsm FI digestion of the 3′ cDNAs and (ii) gel purifying the Bsm FI-released tag fragments before ditag formation. These modifications provide a large quantity of initial 3′ cDNAs and high-quality tags and ditags for the construction of SAGE libraries.