Normalizing and amplifying RNA

US6982143B1 · kind B1 · utility

Assignee

• The Regents of the University of California · US

Inventors

• John Ngai · Boxborough, US
• David M. Lin · Ithaca, US

Key dates

• Filing date: Jun 20, 2000
• Grant date: Jan 3, 2006
• Expiry date: May 15, 2023

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N15/1096
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

The invention provides methods and compositions for normalizing and amplifying RNA populations. The methods generally comprise the steps of copying message RNA (mRNA) to form first single-stranded (ss) cDNA; converting the first ss-cDNA to first double-stranded (ds) cDNA; linearly amplifying the first ds-cDNA to form first amplified RNA (aRNA); tagging the 3′ end of the first aRNA with a known sequence to form 3′-tagged first aRNA; copying the 3′-tagged first aRNA to form second ss-cDNA; and normalizing the mRNA and/or the first aRNA.