Sequencing methods using enzyme conformation

US8911972B2 · kind B2 · utility

Assignee

• PACIFIC BIOSCIENCES OF CALIFORNIA, INC. · US

Inventors

• Mark Chaisson · San Francisco, US
• Sonya Clark · Oakland, US
• Christopher N. SILK · Redwood City, US
• John Lyle · Fremont, US
• Lei Jia · Palo Alto, US
• Keith Bjornson · Fremont, US

Key dates

• Filing date: Dec 14, 2010
• Grant date: Dec 16, 2014
• Expiry date: Sep 11, 2032

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Q1/6869
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Systems and methods are provided for single-molecule sequencing of template nucleic acids in which the signal from a label attached to a polymerase enzyme is used to monitor conformational changes in the polymerase which occur while labeled nucleotides or nucleotide analogs are added to a growing nucleic acid chain which is complementary to the template nucleic acid. The signal indicative of the conformational state of the enzyme is used to determine with higher confidence when true nucleotide or nucleotide analog incorporation events occur, allowing for the improved quality of base calls and sequence determination.