Pacific Biosciences of California, Inc.

Incyte Genomics, Inc.

Walter H Lee

Erik Miller

Satwik Kamtekar

Lei Jia

Arek Bibillo

Pranav Patel

Robin A Emig

Colleen Cutcliffe

Insil Park

Keith P Bjornson

Jeremiah Hanes

Fred C Christians

Glenn K Fu

Laura L Stuve

Molly Min He

Jonathan T Wang

Tyson Bowen

Irene Ni

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties can include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

Recombinant polymerases for improved single molecule sequencing

Provided are compositions comprising recombinant DNA polymerases that include amino acid substitutions, insertions, deletions, and/or exogenous features that confer modified properties upon the polymerase for enhanced single molecule sequencing. Such properties include increased resistance to photodamage, and can also include enhanced metal ion coordination, reduced exonuclease activity, reduced reaction rates at one or more steps of the polymerase kinetic cycle, decreased branching fraction, altered cofactor selectivity, increased yield, increased thermostability, increased accuracy, increased speed, increased readlength, and the like. Also provided are nucleic acids which encode the polymerases with the aforementioned phenotypes, as well as methods of using such polymerases to make a DNA or to sequence a DNA template.

Recombinant polymerases with increased phototolerance

Methods for adding sequence tags during amplification of a nucleic acid target are provided. The target is amplified using two sets of primers that are modified such that only tagged targets can ligate to hairpin adapters that protect them from subsequent exonuclease treatment. Methods of making circular nucleic acids are also provided. Specific overhangs are created by exonuclease trimming with a polymerase such that only desired targets can ligate to sticky ended hairpin adapters that protect them from subsequent exonuclease treatment. Compositions, kits, and systems related to or useful in the methods are also described.

Methods for selectively amplifying and tagging nucleic acids

The present invention provides new methods of synthesizing cDNAs, methods of verifying full-length cDNAs, methods of producing cDNA libraries enriched for full-length inserts, and the like.

cDNA libraries and methods for their production

The present invention provides methods for determining nucleic acid sequences based on known sequences adjacent to the unknown sequences, and reagents for using such methods.

Method for isolating and characterizing nucleic acid sequences

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Campbell, CA, United States of America

Walter H Lee