Illumina Cambridge Limited

Illumina Corporation

The United States of America, As Represented by the Secretary, Department of Health and Human Services

Geoffrey Paul Smith

Roberto Rigatti

Jonathan Mark Boutell

Niall Anthony Gormley

Xiaohai Liu

Xiaolin Wu

Tobias William Barr Ost

David James Earnshaw

David Bentley

Colin Lloyd Barnes

Graham John Worsley

Raquel Maria Sanches-Kuiper

Shujun Luo

David Mark Dunstan Bailey

John Milton

Harold Philip Swerdlow

Anthony Romieu

Gerardo Turcatti

Helmy A Eltoukhy

Robert C Kain

Wenyi Feng

Min-Jui Richard Shen

Bernard Hirschbein

Vincent Peter Smith

Randall Smith

Eric Hans Vermaas

Kay Klausing

Thomas J Joseph

Carolyn Tregidgo

Shankar Balasubramanian

Isabelle Rasolonjatovo

Isabelle Marie Julia Rasolonjatovo

Raquel Maria Sanches

Thomas Joseph

Bernard Moss

Elena Cressina

Michael Mackett

Wouter Meuleman

Rosamond Jackson

Hardold Swerdlow

Liu Xiaohai

Isabelle Marie J Rasolonjatovo

The present invention relates to a method for preparing a library of template polynucleotides and use thereof in methods of solid-phase nucleic acid amplification. More specifically, the invention relates to a method for preparing a library of template polynucleotides that have common sequences at their 5' ends and at their 3′ ends.

Method of preparing libraries of template polynucleotides

Presented are methods and compositions for using immobilized transposase and a transposon end for generating an immobilized library of 5′-tagged double-stranded target DNA on a surface. The methods are useful for generating 5′- and 3′-tagged DNA fragments for use in a variety of processes, including massively parallel DNA sequencing.

Sample preparation on a solid support

The present disclosure provides methods and systems for detecting multiple different nucleotides in a sample. In particular, the disclosure provides for detection of multiple different nucleotides in a sample utilizing fewer detection moieties than the number of nucleotides being detected and/or fewer imaging events than the number of nucleotides being detected.

Methods and compositions for nucleic acid sequencing

The invention relates to methods for pairwise sequencing of a double-stranded polynucleotide template, which permit the sequential determination of nucleotide sequences in two distinct and separate regions on complementary strands of the double-stranded polynucleotide template. The two regions for sequence determination may or may not be complementary to each other.

Method for sequencing a polynucleotide template

The invention relates to modified polymerase enzymes which exhibit improved incorporation of nucleotide analogues bearing substituents at the 3' position of the sugar moiety that are larger in size than the naturally occurring 3&#x2032; hydroxyl group. Also described are methods of using the polymerases to incorporate nucleotides into polynucleotides, particularly in the context of DNA sequencing.

Modified polymerases for improved incorporation of nucleotide analogues

The present invention relates to a method for preparing a library of template polynucleotides and use thereof in methods of solid-phase nucleic acid amplification. More specifically, the invention relates to a method for preparing a library of template polynucleotides that have common sequences at their 5' ends and at their 3&#x2032; ends.

Presented are methods and compositions for using immobilized transposase and a transposon end for generating an immobilized library of 5'-tagged double-stranded target DNA on a surface. The methods are useful for generating 5&#x2032;- and 3&#x2032;-tagged DNA fragments for use in a variety of processes, including massively parallel DNA sequencing.

Some embodiments described herein relate to modified nucleotide and nucleoside molecules with novel 3'-hydroxy protecting groups. Also provided herein are methods to prepare such modified nucleotide and nucleoside molecules and sequencing by synthesis processes using such modified nucleotide and nucleoside molecules.

Modified nucleosides or nucleotides

The invention provides methods for pairwise sequencing of a double-stranded polynucleotide template, which methods result in the sequential determination of nucleotide sequences in two distinct and separate regions of the polynucleotide template.

The invention relates to methods of generating templates for a nucleic acid sequencing reaction which comprise: providing at least one double-stranded nucleic acid molecule, wherein both strands of the double-stranded nucleic acid molecule are attached to a solid support at the 5' end, cleaving one or both strands of the double-stranded nucleic acid molecule, and subjecting the cleaved strand(s) to denaturing conditions to remove the portion of the cleaved strand(s) not attached to the solid support, thereby generating a partially or substantially single-stranded template for a nucleic acid sequencing reaction.

Preparation of templates for nucleic acid sequencing

The invention relates to methods for pairwise sequencing of a double stranded polynucleotide template, which permit the sequential determination of nucleotide sequences in two distinct and separate regions on complementary strands of the double-stranded polynucleotide template. The two regions for sequence determination may or may not be complementary to each other.

A reaction mixture including (a) a nucleic acid having a primer hybridized to a template, (b) nucleotide analogs, wherein the nucleotide analogs have a blocking moiety; (c) a polymerase that is capable of forming an extended primer by adding the nucleotide analogs to the primer, and (d) a deblocking agent that is capable of removing the blocking moiety from the extended primer. Also provided is a method of synthesizing a polynucleotide including sequentially adding a plurality of the different nucleotides analogs to the nucleic acid via several reaction cycles in the reaction mixture, wherein each reaction cycle includes (i) the polymerase adding a nucleotide analog to the nucleic acid to form a transient nucleic acid species comprising a blocking moiety, and (ii) the deblocking agent modifying the transient nucleic acid species to remove the blocking moiety.

Continuous extension and deblocking in reactions for nucleic acids synthesis and sequencing

Growing community of inventors

Nr Saffron Walden, United Kingdom

Geoffrey Paul Smith