Illumina Cambridge Limited

Wouter Meuleman

Rosamond Jackson

Kevin L Gunderson

Xiaohai Liu

Molly Min He

Niall Anthony Gormley

Geoffrey Paul Smith

Frank J Steemers

Amirali Kia

Igor Yu Goryshin

Jason Richard Betley

Rigo Pantoja

Dmitry K Pokholok

Elena Cressina

Natalie Morrell

Fan Zhang

Avgousta Ioannou

Gareth Jenkins

Jacqueline Weir

Steven J Norberg

Embodiments provided herein relate to methods and compositions for preparing an immobilized library of barcoded DNA fragments of a target nucleic acid, identifying genomic variants, determining the contiguity information, phasing information, and methylation status of the target nucleic acid.

Contiguity preserving transposition

A method for determining the sequence of a target nucleic acid, including steps of contacting a target nucleic acid with a polymerase to sequentially remove nucleotide triphosphates from the target nucleic acid, wherein the nucleotide triphosphates that are removed have a variety of different base moieties; and distinguishing the different base moieties for the nucleotide triphosphates that are removed. Also provided is a apparatus including a nanopore positioned in a fluid impermeable barrier to form a passage through which a nucleotide triphosphate can pass from a first fluid reservoir to a second fluid reservoir, and a reaction mix in the first fluid reservoir that includes a polymerase, target nucleic acid having two strands, and pyrophosphorolytic concentration of pyrophosphate.

Pyrophosphorolytic sequencing

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

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Nr Saffron Walden, United Kingdom

Wouter Meuleman