Roche Sequencing Solutions, Inc.

Roche Molecular Systems, Inc.

Genia Technologies, Inc.

Preethi Sarvabhowman

Aruna Ayer

Eileen Thai

Ilya Lederman

Dhruti Vasudev Dalal

Arkadiusz Bibillo

Charles Wayan Schwab

Shawn Lee Suko

Mara Boenitz-Dulat

Cleoma Arnold

Colin Alexander McGaw

Barbara Eckert

David Daniel Wunderlich

Bigna Woersdoerfer

Evan Amato

Described herein are polymerase variants that are exonuclease deficient. Some variants retain the strand displacement capability comparable to the wild-type or parental polymerase. Some variants have a strand displacement capability that is improved relative to the wild-type or parental polymerase. The variants may have an extension rate that is greater than the wild-type or parental polymerase. The variants may have a waiting time that is less than the wild-type or parental polymerase.

Exonuclease deficient polymerases

The present disclosure provides methods and compositions for enhancing the processivity of a polymerase in catalyzing template-dependent DNA synthesis in high concentrations of salt. Also disclosed are methods and compositions for enhancing the assembly of polymerase-template complex compatible with active DNA synthesis in the presence of low levels of nucleotides and at a high temperature, such as temperatures at or near the melting temperature of the polymerase.

Polymerase-template complexes

The present disclosure provides variant Pol6 polymerase polypeptides, compositions comprising the Pol6 variant polypeptides, and methods for using the variant Pol6 polypeptides for determining the sequencing of nucleic acids, for example, by nanopore sequencing. The variant Pol6 polymerases possess decreased rates of dissociation of template from the polymerase-template complex, which result in increased processivity relative to the parental Pol6 polypeptides from which they are derived.

Pol6 polymerase variants

Polymerase variants

The present of disclosure provides variant Pol6 polymerase polypeptide, compositions comprising the Pol6 variant polypeptides, and methods for using the variant Pol6 polypeptides for determining the sequencing of nucleic acids, for example, by nanopore sequencing. The variant Pol6 polymerases possess decreased rates of dissociation of template from the polymerase-template complex, which result in increased processivity relative to the parental Pol6 polypeptides from which they are derived.

Growing community of inventors

Santa Clara, CA, United States of America

Preethi Sarvabhowman