Microarray Hybridization with Fractionated cDNA: Enhanced Identification of Differentially Expressed Genes

✍ Scribed by Kohji Sakai; Hirofumi Higuchi; Kenichi Matsubara; Kikuya Kato

• Book ID: 102565778
• Publisher: Elsevier Science
• Year: 2000
• Tongue: English
• Weight: 141 KB
• Volume: 287
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.2000.4831

✦ Synopsis

Molecular indexing is a technique to select a subpopulation of cDNA by ligation of adapters to cDNA fragments digested by a class IIS restriction enzyme(s). By this technique, 3' end cDNA fragments are divided into 16 fractions by selective ligation of adapters and subsequent PCR amplification. Each fraction is used as a hybridization target for microarray hybridization. This fractionated target has a relatively lower nucleic acid complexity, including more fractions for rare transcripts, and is useful for their detection. In an experiment with mouse cerebellum and cerebrum RNA, analysis of a subset of candidate genes indicated that 10 times more differentially expressed genes were detected by this method than by the conventional method using the unfractionated target.