Illumina Cambridge Limited

Solexa, Inc.

Illumina Corporation

Colin Lloyd Barnes

Xiaohai Liu

John Milton

Xiaolin Wu

Shankar Balasubramanian

Roberto Rigatti

Geoffrey Paul Smith

Tobias William Barr Ost

Graham John Worsley

Harold Philip Swerdlow

Silke Ruediger

Joseph Brennan

Mark Smith

Gerardo Turcatti

Jonathan Mark Boutell

Niall Anthony Gormley

Eric Hans Vermaas

Anthony Romieu

David James Earnshaw

Vincent Peter Smith

David Bentley

Isabelle Rasolonjatovo

Nikolai Nikolaevich Romanov

Isabelle Marie Julia Rasolonjatovo

Dale Buermann

Mark T Reed

Mark R Pratt

Jason Bryant

John Bridgham

Klaus Maisinger

David Klenerman

Bojan Obradovic

Mark Allen Osborne

Gary Schroth

Saibal Banerjee

Kevin Benson

Sergey Etchin

Johnny Ho

Xavier Lee

Peter Lundberg

Chiara Rodighiero

Subra Sankar

Darren James Ellis

Tom Brown

John Andrew Todd

Richard Michael Durbin

Liu Xiaohai

Isabelle Marie J Rasolonjatovo

Jonny Ho

Ning Sizto

Gerado Turcatti

Novel rhodamine dye compounds, labelled conjugates comprising the dyes are described, together with methods for their use. The dyes and labelled conjugates are useful as molecular probes in a variety of applications, such as in assays involving staining of cells, protein binding, and analysis of nucleic acids, such as hybridization assays and nucleic acid sequencing.

Dye compounds and the use of their labelled conjugates

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.

Method for sequencing a 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

Nucleosides and nucleotides are disclosed that are linked to detectable labels via a cleavable linker group.

Labelled nucleotides

The invention provides modified nucleotide or nucleoside molecule comprising a purine or pyrimidine base and a ribose or deoxyribose sugar moiety having a removable 3'-OH blocking group covalently attached thereto, such that the 3&#x2032; carbon atom has attached a group of the structure &#x2014;O&#x2014;Z wherein Z is any of &#x2014;C(R&#x2032;)2-O&#x2014;R&#x2033;, &#x2014;C(R&#x2032;)2-N(R&#x2033;)2, &#x2014;C(R&#x2032;)2-N(H)R&#x2033;, &#x2014;C(R&#x2032;)2-S&#x2014;R&#x2033; and &#x2014;C(R&#x2032;)2-F, wherein each R&#x2033; is or is part of a removable protecting group; each R&#x2032; is independently a hydrogen atom, an alkyl, substituted alkyl, arylalkyl, alkenyl, alkynyl, aryl, heteroaryl, heterocyclic, acyl, cyano, alkoxy, aryloxy, heteroaryloxy or amido group, or a detectable label attached through a linking group; or (R&#x2032;)2 represents an alkylidene group of formula &#x2550;C(R&#x2032;&#x2033;)2 wherein each R&#x2032;&#x2033; may be the same or different and is selected from the group comprising hydrogen and halogen atoms and alkyl groups; and wherein said molecule may be reacted to yield an intermediate in which each R&#x2033; is exchanged for H or, where Z is &#x2014;C(R&#x2032;)2-F, the F is exchanged for OH, SH or NH2, preferably OH, which intermediate dissociates under aqueous conditions to afford a molecule with a free 3&#x2032;OH; with the proviso that where Z is &#x2014;C(R&#x2032;)2-S&#x2014;R&#x2033;, both R&#x2032; groups are not H.

Modified nucleotides

Method for sequencing a polynucelotide template

Growing community of inventors

Cambridge, United Kingdom

Colin Lloyd Barnes