Generation of modified polymerases for improved accuracy in single molecule sequencing

US8257954B2 · kind B2 · utility

Assignee

• PACIFIC BIOSCIENCES OF CALIFORNIA, INC. · US

Inventors

• Sonya Clark · Oakland, US
• Arek Bibillo · Cupertino, US
• Paul Peluso · East Palo Alto, US
• Fred Christians · Los Altos Hills, US
• Molly He · San Diego, US
• Insil Park · Fremont, US
• Harold Lee · Holly Springs, US
• Keith Bjornson · Fremont, US
• Lei Jia · Palo Alto, US
• Robin Emig · Belmont, US

Key dates

• Filing date: Mar 30, 2009
• Grant date: Sep 4, 2012
• Expiry date: May 24, 2030

Classification

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

Abstract

Provided are compositions comprising modified recombinant polymerases that exhibit branching fractions that are less than the branching fractions of the polymerases from which they were derived, or branching fractions that are less than about 25% for a phosphate-labeled nucleotide analog. Also provided are compositions comprising modified recombinant polymerases that exhibit closed polymerase/DNA complexes with increased stability relative to the parental polymerases. Also provided are compositions comprising modified recombinant polymerases that exhibit decreased rate constants relative to the parental polymerases. Provided are methods for generating polymerases with the aforementioned phenotypes. Provided are methods of using such polymerases to make a DNA or to sequence a DNA template.