Rolling circle amplification detection of RNA and DNA

US6783943B2 · kind B2 · utility

Assignee

• The Regents of the University of California · US

Inventors

• Allen T. Christian · Wildwood, US
• Melissa S. Pattee · Livermore, US
• Cristina M. Attix · Livermore, US
• James Tucker · Rochester, US

Key dates

• Filing date: Dec 18, 2001
• Grant date: Aug 31, 2004
• Expiry date: Apr 1, 2022

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6806
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Rolling circle amplification (RCA) has been useful for detecting point mutations in isolated nucleic acids, but its application in cytological preparations has been problematic. By pretreating cells with a combination of restriction enzymes and exonucleases, we demonstrate RCA in solution and in situ to detect gene copy number and single base mutations. It can also detect and quantify transcribed RNA in individual cells, making it a versatile tool for cell-based assays.