Element Biosciences, Inc.

Illumina Corporation

Illumina Cambridge Limited

Samantha Snow

Junhua Zhao

William Light

Michael Previte

Molly Min He

Matthew William Kellinger

Sinan Arslan

Hua Yu

Yu-Hsien Hwang-Fu

Michael Kim

Marco Tjioe

Andrew Boddicker

Grace DeSantis

Andrew Slatter

Natalie Morrell

Jian-Sen Li

Stephen M Gross

Kevin Shen

Tyler Lopez

Mark Ambroso

Virginia Saade

Michael Klein

The present disclosure provides compositions comprising nucleic acid double-stranded splint adaptors, including kits, and methods that employ the double-stranded splint adaptors. The double-stranded splint adaptors () can be used in a one-pot, multi-enzyme reaction to introduce one or more new adaptor sequences into a library molecule. The double-stranded splint adaptor () comprises a first splint strand (long splint strand ()) and a second splint strand (short splint strand ()), where the first and second splint strands are hybridized together to form the double-stranded splint adaptor () having a double-stranded region and two flanking single-stranded regions. The second splint strand () carries the new adaptor sequence(s) to be introduced, such as for example a universal binding sequence and/or an index sequence.

Double-stranded splint adaptors and methods of use

The present disclosure relates to methods, compositions, and kits for treating target nucleic acids, including methods and compositions for fragmenting and tagging nucleic acid (e.g., DNA) using transposome complexes bound to a solid support.

Tagmentation using immobilized transposomes with linkers

The present disclosure provides compositions and methods that employ the compositions for conducting pairwise sequencing and for generating concatemer template molecules for pairwise sequencing. The concatemers can be generated using a rolling circle amplification reaction which is conducted either on-support, or conducted in-solution and then distributed onto a support. The rolling circle amplification reaction generates concatemers containing tandem copies of a sequence of interest and at least one universal adaptor sequence. An increase in the number of tandem copies in a given concatemer increases the number of sites along the concatemer for hybridizing to multiple sequencing primers which serve as multiple initiation sites for polymerase-catalyzed sequencing reactions. When the sequencing reaction employs detectably labeled nucleotides and/or detectably labeled multivalent molecules (e.g., having nucleotide units), the signals emitted by the nucleotides or nucleotide units that participate in the parallel sequencing reactions along the concatemer yields an increased signal intensity for each concatemer.

Compositions and methods for pairwise sequencing

Growing community of inventors

San Diego, CA, United States of America

Samantha Snow