Roche Sequencing Solutions, Inc.

Roche Diagnostics Gmbh

F. Hoffmann-la Roche Ag

Peter Crisalli

Aruna Ayer

Meng Taing

Frank Bergmann

Dmitriy Gremyachinskiy

Randall W Davis

Helen Franklin

Joshua N Mabry

Omid Khakshoor

Natalie B Chechelski Johnston

Ashwini Bhat

Dieter Heindl

Christoph Seidel

Arkadiusz Bibillo

Cynthia Ann Cech

Timothy Kellogg Craig

Kirti Dhiman

Andrew Trans

Hannes Kuchelmeister

Sebastian Johannes Pomplun

Christian Wellner

Ian Quang Chau

Sara Kelley

Dieter Hiendl

Hasham Saqib

Hannah Marie Kallewaard-Lum

Lars Hillringhaus

Seong-Ho Shin

Cynthia Cech

Rongxin Nie

The disclosure relates to tagged nucleoside compounds comprising a nucleotide polyphosphate covalently attached to a tag, wherein the compound is a polymerase substrate and the polymer moiety is capable of entering a nanopore linked to the polymerase and thereby altering the flow of ions through the nanopore. The disclosure also provides methods for preparing the tagged nucleoside compounds and for their use as tags in nanopore-based nucleic acid detection and sequencing.

Tagged nucleoside compounds useful for nanopore detection

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 3' protected nucleotides, including those 3&#x2032; protected nucleotides having a detectable tag. Systems and methods of sequencing nucleic acids using the 3&#x2032; protected nucleotides are also disclosed, such as the sequencing of a nucleic acid using a nanopore or the sequencing of a nucleic acid via sequencing-by-synthesis.

3' protected nucleotides

The present invention relates to improving the processing rate of a sequencing reaction, for example in a nanopore sequencing reaction, by means of using improved nucleoside-tags. The tags are linked to the nucleoside phosphate via a Pictet Spengler reaction. Exemplary sequencing reactions that are improved by the present methods include nanopore-based nucleic acid sequencing-by-synthesis reactions.

Method for labeling of aldehyde containing target molecules

The present disclosure relates to compounds comprising a negatively-charged polymer moiety which is capable of entering a nanopore and upon entering a nanopore in the presence of positive ions results in an increased flow of the positive ions through the nanopore. The present disclosure provides methods of preparing the compounds and for their use as nanopore-detectable tags, in particular, for nanopore-based nucleic acid detection and sequencing.

Tagged nucleotides useful for nanopore detection

The present disclosure relates to tagged multi-nucleotide compounds, which comprise a single tag moiety covalently linked to a plurality of nucleoside-5'-oligophosphate moieties. As disclosed herein, these tagged multi-nucleotide compounds have improved characteristics as polymerase substrates and can be used in a range of nucleic acid detection and sequencing methods, including nanopore sequencing-by-synthesis.

Tagged multi-nucleotides useful for nucleic acid sequencing

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Mountain View, CA, United States of America

Peter Crisalli