Linear amplification mediated PCR (LAM PCR)

US6514706B1 · kind B1 · utility

Inventors

• Christoph Von Kalle · Schallstadt, DE
• Manfred Schmidt · Krefeld, DE

Key dates

• Filing date: Jul 16, 2001
• Grant date: Feb 4, 2003
• Expiry date: Jul 16, 2021

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6855
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A high sensitive method for identifying and/or sequencing an unknown DNA or RNA sequence flanking a known DNA or RNA region is described, wherein,(a) one or more DNA or RNA fragments are subjected in a first step to one or more linear PCR steps using one more primers,(b) the single strands obtained are complemented to form double strands,(c) the double strands are digested by one or more restriction enzymes in order to produce smooth and/or cohesive ends,(d) an oligonucleotide of known sequence is added at the digested ends, and(e) the thus obtained DNA fragments are amplified and detected.