Pacific Biosciences of California, Inc.

Lei Sun

Natasha Popovich

Stephen Wilson Turner

Sassan Sheikholeslami

Gene W Shen

Jonas Korlach

Steven Lin

Jaime Juan Benitez-Marzan

Erik Miller

Michael W Hunkapiller

Satwik Kamtekar

Thang Pham

Aparna Vedula

Stephen T Yue

John Lyle

Andrei Fedorov

Benjamin A Flusberg

Jeremy Gray

Insil Park

David Christopher Scherer

Robin A Emig

John S Eid

Fred C Christians

Adrian Nielsen Fehr

Stephen P Dudek

Robert Sebra

Ravi Dalal

Yufeng Qian

Methods, compositions, and systems for distributing nucleic acids into array regions are provided. The methods, compositions, and systems utilize nucleic acid condensing agents to increase efficiency of distribution of the nucleic acids into the array regions. Various methods for facilitating distribution of the nucleic acids to the array regions are provided.

Loading nucleic acids onto substrates

The present disclosure provides improved methods for isolating asymmetrically-primed and/or asymmetrically-tagged nucleic acid complexes that find use in downstream analytical analyses, including sequence analysis. Compositions comprising such complexes and kits and systems for generating such complexes are also provided.

Methods and compositions for isolating asymmetric nucleic acid complexes

The present disclosure provides methods, compositions, and systems for distributing polymerase compositions into array regions. In particular, the described methods, compositions, and systems utilize density differentials and/or additives to increase efficiency in the distribution of polymerase compositions to a surface as compared to methods utilizing only diffusion control.

Methods and compositions for loading of polymerase complexes

The present invention provides methods, compositions, and systems for distributing molecules and complexes into reaction sites. In particular, the methods, compositions, and systems of the present invention result in loading of polymerase enzyme complexes into a predetermined number of reaction sites, including nanoscale wells.

Methods and compositions for delivery of molecules and complexes to reaction sites

Real time redox sequencing methods, devices, and systems are described. Arrays of redox devices comprising one or two electrodes are used to provide sequence information about a template nucleic acid in a polymerase-template complex bound proximate to the electrode(s). A sequencing reaction mixture comprising nucleotide analogs comprising redox labels is introduced to the array of redox devices under conditions of polymerase mediated nucleic acid synthesis. The time sequence of incorporation of nucleotide analogs is determined by electrochemically identifying the redox labels of the nucleotide analogs that are incorporated into the growing strand.

Real-time redox sequencing chips

The application relates to methods of analyzing luminescent species. A substrate is provided that has a plurality of zero mode waveguides having apertures that extend through an upper non-reflective layer that is disposed on a lower transparent layer of a substrate. The apertures have non-reflective oxide layers on the reflective side walls of the apertures, the side walls having a thickness of greater than 10 nm, and the oxide layer is formed by oxidizing the non-reflective layer. The volume within the oxide layer defines a solution volume, and the volume within the reflective walls defines a ZMW volume. Having such non-reflective layers on the walls of the ZMW usefully decouples the solution volume from the ZMW volume.

Method for analyzing luminescent species

The present invention provides methods, compositions, and systems for distributing single polymerase molecules into array regions. In particular, the methods, compositions, and systems of the present invention result in a distribution of single polymerase molecules into array regions at a percentage that is larger than the percentage expected to be occupied under a Poisson distribution.

Methods and compositions for single molecule composition loading

Real-time redox sequencing methods

Methods, Compositions, and Systems are provided for obtaining polymerase-template complex mixtures with improved levels of active polymerase. In some aspects active polymerase-template complex is separated by affinity for nucleoside phosphate moieties attached to a resin. In some aspects, a polymerase-template complex is exposed to reaction conditions in which a complementary strand to the template is produced. The extended reaction mixture is purified by reaction with a resin comprising nucleoside phosphate moieties. This purified mixture can be loaded onto substrates and can be used for further analyzes including single molecule sequencing.

Purified polymerase / template complexes

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San Jose, CA, United States of America

Lei Sun