Generation of high-quantity and quality tag/ditag cDNAs for SAGE analysis

S. Lee, J. Chen, G. Zhou, S. M. Wang

Research output: Contribution to journalArticle

25 Scopus citations

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 BsmFI digestion of the 3′ cDNAs and (ii) gel purifying the BsmFI-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.

Original languageEnglish (US)
Pages (from-to)348-354
Number of pages7
JournalBioTechniques
Volume31
Issue number2
StatePublished - Aug 25 2001
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

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    Lee, S., Chen, J., Zhou, G., & Wang, S. M. (2001). Generation of high-quantity and quality tag/ditag cDNAs for SAGE analysis. BioTechniques, 31(2), 348-354.