Recombinant polymerases for improved single molecule sequencing

US8999676B2 · kind B2 · utility

Assignee

• PACIFIC BIOSCIENCES OF CALIFORNIA, INC. · US

Inventors

• Robin Emig · Belmont, US
• Lei Jia · Palo Alto, US
• Satwik Kamtekar · Mountain View, US
• Erik Miller · Seattle, US
• Colleen Cutcliffe · Menlo Park, US
• Walter Lee · Campbell, US

Key dates

• Filing date: Jul 5, 2011
• Grant date: Apr 7, 2015
• Expiry date: Oct 29, 2032

Classification

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

Abstract

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.