Roche Sequencing Solutions, Inc.

Kapa Biosystems, Inc.

Roche Diagnostics Gmbh

F. Hoffmann-la Roche Ag

Severine Margeridon

Martin Ranik

Ashley Hayes

Richard Dannebaum

Daleen Badenhorst

Jenny A Johnson

Rui Chen

Alexandra Hui Wang

Monica Herrera

Persis Wadia

Aruna Arcot

Toummy Guettouche

Michael S Berry

Etienne Slabbert

Michael Berry

The invention is a novel method of sequencing nucleic acids involving making and sequencing a library of single stranded circular target nucleic acids.

Generation of single-stranded circular DNA templates for single molecule sequencing

The invention is a novel method of separately sequencing each strand of a nucleic acid involving the use of an adaptor comprising a strand cleavage site or a strand synthesis termination site. The adaptor may also be self-priming at the strand cleavage site.

The invention is a novel method of making and using a library such as a sequencing library of single stranded circular nucleic acid templates via splint ligation. In particular, disclosed are methods of making circular target nucleic acid molecules and libraries of such molecules for downstream analysis such as nucleic acid sequencing. The method comprises the steps of adding universal sequences to nucleic acid molecules, rendering single-stranded these nucleic acid molecules with universal sequences on their ends by contacting with a probe complementary to at least a portion of the universal sequences, and allowing the hybridized probe to enable circularization and formation of single-stranded circular (sscDNA) molecules.

Hybridization-extension-ligation strategy for generating circular single-stranded DNA libraries

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Castro Valley, CA, United States of America

Severine Margeridon