Illumina Cambridge Limited

Rosamond Jackson

Niall Anthony Gormley

Natalie Morrell

Wouter Meuleman

Avgousta Ioannou

Kevin L Gunderson

Xiaohai Liu

Molly Min He

Geoffrey Paul Smith

Frank J Steemers

Amirali Kia

Igor Yu Goryshin

Jason Richard Betley

Rigo Pantoja

Dmitry K Pokholok

Elena Cressina

Fan Zhang

Gareth Jenkins

Jacqueline Weir

Steven J Norberg

Igor Goryshin

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

Presented herein are methods and compositions surface-based tagmentation. In particular embodiments, methods of preparing an immobilized library of fragmented and tagged DNA molecules on a solid surface are presented. In particular embodiments, the solid surface comprises immobilized transposomes in a dried format, suitable for reconstitution upon contact with liquid, such as a liquid sample.

Surface-based tagmentation

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

Rosamond Jackson