Bio-rad Laboratories, Inc.

Roche Sequencing Solutions, Inc.

Genia Technologies, Inc.

Roche Molecular Systems, Inc.

Pacific Biosciences of California, Inc.

Roche Diagnostics Gmbh

F. Hoffmann-la Roche Ag

Arkadiusz Bibillo

Pranav Patel

Aruna Ayer

Randall W Davis

Eileen Thai

Ilya Lederman

Roger Chen

Shawn Lee Suko

Mara Boenitz-Dulat

Cynthia Ann Cech

Timothy Kellogg Craig

Peter Crisalli

Helen Franklin

Charles Wayan Schwab

Colin Alexander McGaw

Dhruti Vasudev Dalal

Cleoma Arnold

Tyler O'Brien Shultz

Barbara Eckert

Daniel Korenblum

Preethi Sarvabhowman

David Daniel Wunderlich

Bigna Woersdoerfer

Kirti Dhiman

Natalie B Chechelski Johnston

Joshua N Mabry

Christopher V Reggiardo

Jonathan Petersen

Jonas Korlach

Roger J A Chen

Keith P Bjornson

Jeremiah Hanes

Michael Dorwart

Kevin Deierling

Cynthia Cech

Shawn Suko

Cleoma Renetta Arnold

Tyler O'Brien Schultz

Methods, systems, and devices for sampling/isolating material from cells. An exemplary system may comprise a chip including an electrode array of sampling electrodes arranged along a surface of the chip. A cell-receiving area may be located adjacent the surface of the chip. The system also may comprise a tag array of tags supported by the chip and aligned with the electrode array. Each tag of the tag array may include an identifier that is unique to the tag within the tag array. Each tag may be configured to bind nucleic acids, or a capturing agent distinct from the tag may be aligned with each sampling electrode of the electrode array to capture a protein or other analyte of interest. The system further may comprise a control circuit configured to apply an individually controllable voltage to each sampling electrode of the electrode array and measure an electrical property of the sampling electrode.

Method and system for sampling material from cells

The present disclosure provides methods and systems for amplifying and analyzing nucleic acid samples. The present disclosure provides methods for preparing cDNA and/or DNA molecules and cDNA and/or DNA libraries using modified reverse transcriptases.

Methods for processing nucleic acid samples

Methods of cDNA preparation

Modified reverse transcriptases

Hybrid reverse transcriptases are provided that comprise a non-retroviral retrotransposon, or a fragment of the non-retroviral retrotransposon having reverse transcriptase activity, joined to a nucleic acid binding protein. Also provided are methods of using the hybrid reverse transcriptases to prepare a cDNA molecule library.

Hybrid reverse transcriptases

Disclosed are methods for isolating polymerase complexes from a mixture of polymerase complex components. The polymerase complexes can comprise a nanopore to provide isolated nanopore sequencing complexes. The methods relate to the positive and negative isolation of the polymerase complexes and/or nanopore sequencing complexes. Also disclosed is a nucleic acid adaptor for isolating active polymerase complexes, polymerase complexes comprising the nucleic acid adaptor, and methods for isolating active polymerase complexes using the nucleic acid adaptor.

Purification of polymerase complexes

The present disclosure provides methods and systems for amplifying and analyzing nucleic acid samples. The present disclosure provides methods for preparing cDNA and/or DNA molecules ad cDNA and/or DNA libraries using modified reverse transcriptases.

Methods for producing modified reverse transcriptases

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

Sequencing adaptors and methods are provided for preparation of polynucleotides for sequencing. The sequencing adaptors contain a portion of a recognition sequence for a methyl-dependent endonuclease. Unwanted adaptor dimers that form during ligation of adaptors to target polynucleotides produce a complete restriction sequence and are cleaved by the endonuclease, followed by exonuclease digestion, thereby removing the dimers.

Methods for removal of adaptor dimers from nucleic acid sequencing preparations

This disclosure provides chips, systems and methods for sequencing a nucleic acid sample. Tagged nucleotides are provided into a reaction chamber comprising a nanopore in a membrane. An individual tagged nucleotide of the tagged nucleotides can contain a tag coupled to a nucleotide, which tag is detectable with the aid of the nanopore. Next, an individual tagged nucleotide of the tagged nucleotides can be incorporated into a growing strand complementary to a single stranded nucleic acid molecule derived from the nucleic acid sample. With the aid of the nanopore, a tag associated with the individual tagged nucleotide can be detected upon incorporation of the individual tagged nucleotide. The tag can be detected with the aid of the nanopore when the tag is released from the nucleotide.

Nucleic acid sequencing using tags

Described herein is a variant pol6 polymerase having at least one mutation selected from H223, N224, Y225, H227, I295, Y342, T343, I357, S360, L361, I363, S365Q, S366, Y367, P368, D417, E475, Y476, F478, K518, H527, T529, M531, N535, G539, P542, N545, Q546, A547, L549, I550, N552, G553, F558, A596, G603, A610, V615, Y622, C623, D624, I628, Y629, R632, N635, M641, A643, I644, T647, I648, T651, I652, K655, W656, D657, V658, H660, F662, L690 and combinations thereof.

Polymerase variants

Growing community of inventors

Cupertino, CA, United States of America

Arkadiusz Bibillo