Work

Generation of High-Quantity and Quality Tag/Ditag cDNAs for SAGE Analysis

Year: 2001

Type: article

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.

Source: BioTechniques

DOI: 10.2144/01312st07

Authors: Sanggyu Lee, Jianjun Chen, Guolin Zhou, San Ming Wang

Institution: University of Chicago Medical Center

Language: English

Topic: Genomics and Phylogenetic Studies

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $1,415

Funders: National Institutes of Health, National Cancer Institute