Pacific Biosciences of California, Inc.

Elan Pharmaceuticals, Inc.

Other

Pharmacia & Upjohn Company

Louis Brogley

Lubomir Sebo

Stephen T Yue

Gene W Shen

Andrei Fedorov

Jeremiah Hanes

Honey Osuna

Yuri Lapin

John Lyle

Frank Zheng

Keith P Bjornson

Erik Miller

Satwik Kamtekar

John Alan Tucker

Eugene D Thorsett

Michael A Pleiss

Francine S Grant

Michael S Dappen

Michael Ye

Jon Eric Hawkinson

Sarah Bartulis

Ramesh Kasar

Varghese John

Martin Neitzel

Wei-Chuan Sun

Mohammed A Khan

David T Connor

Bruce D Roth

Jay S Tung

Darren B Dressen

Michael Francis Rafferty

Harriet W Hamilton

Jeffrey Wegener

Simeon Bowers

Yuntao Song

James Beck

Thomas C Malone

Lain-Yen Hu

Barbara Jagodzinska

Richard John Booth

Wayne Livingston Cody

Shon R Pulley

Gary D Probst

Michel Maillard

Brian Alan Sherer

Laszlo Nadasdi

Jon S Jacobs

Balazs G Szoke

Laszlo Urge

John Xiaoqiang He

Jose Aquino

Ryan C Holcomb

Diego F Silva

Neerav Shah

Ying Zi Xu

Leonard Joseph Lescosky

Labeled nucleotide analogs comprising at least one avidin protein, at least one dye-labeled compound, and at least one nucleotide compound are provided. The analogs are useful in various fluorescence-based analytical methods, including the analysis of highly multiplexed optical reactions in large numbers at high densities, such as single molecule real time nucleic acid sequencing reactions. The analogs are detectable with high sensitivity at desirable wavelengths. They contain structural components that modulate the interactions of the analogs with DNA polymerase, thus decreasing photodamage and improving the kinetic and other properties of the analogs in sequencing reactions. Also provided are nucleotide and dye-labeled compounds of the subject analogs, as well as intermediates useful in the preparation of the compounds and analogs. Compositions comprising the compounds, methods of synthesis of the intermediates, compounds, and analogs, and mutant DNA polymerases are also provided.

Labeled nucleotide analogs, reaction mixtures, and methods and systems for sequencing

Compositions, methods, and systems are provided for fluorescent polymerase enzyme substrates comprising protein shields for improving enzyme photostability in single molecule real time sequencing. Fluorescent polymerase enzyme substrates of the invention have a protein shield between the fluorescent dye moieties and nucleotide moieties of the polymerase enzyme substrate. The polymerase enzyme substrates have a nucleotide component and a dye component, each attached to a protein. The attachments can be covalent. The protein can, for example, prevent the direct interaction of the fluorescent dye moiety with the enzyme when carrying out nucleotide synthesis, preventing photodamage to the enzyme. The polymerase enzyme substrates of the invention can have multiple dyes and multiple nucleotide moieties.

Fluorescent polymerase enzyme substrates having protein shields

Protected fluorescent reagent compounds and their methods of synthesis are provided. The compounds are useful in various fluorescence-based analytical methods, including the analysis of highly multiplexed optical reactions in large numbers at high densities, such as single molecule real time nucleic acid sequencing reactions. The compounds contain fluorescent dye elements, that allow the compounds to be detected with high sensitivity at desirable wavelengths, binding elements, that allow the compounds to be recognized specifically by target biomolecules, and protective shield elements, that decrease undesirable contacts between the fluorescent dye elements and the bound target biomolecules and that therefore decrease photodamage of the bound target biomolecules by the fluorescent dye elements.

Protected fluorescent reagent compounds

Modified nucleotide reagents

Multimeric protected fluorescent reagents and their methods of synthesis are provided. The reagents are useful in various fluorescence-based analytical methods, including the analysis of highly multiplexed optical reactions in large numbers at high densities, such as single molecule real time nucleic acid sequencing reactions. The reagents contain fluorescent dye elements, that allow the compounds to be detected with high sensitivity at desirable wavelengths, binding elements, that allow the compounds to be recognized specifically by target biomolecules, and protective shield elements, that decrease undesirable contacts between the fluorescent dye elements and the bound target biomolecules and that therefore decrease photodamage of the bound target biomolecules by the fluorescent dye elements. The reagents also contain coupling elements connect monomeric compounds into multimeric forms, thereby increasing brightness.

Multimeric protected fluorescent reagents

Growing community of inventors

Santa Cruz, CA, United States of America

Louis Brogley