Other

The Perkin-elmer Corporation

California Institute of Technology

Applied Biosystems, LLC

John H Richards

Michael W Hunkapiller

Robert Austin Stringer

Charles S Whipple, Jr

John Brandis

Curtis Bloom

Sheila A Iverson

Ken Maloney

Dianne M Perez

The present teachings relate to DNA polymerases that have an F667Y mutation as defined with respect to Taq DNA polymerase and exhibit reduced discrimination against labeled nucleotides into polynucleotides. The teachings also include kits comprising the subject DNA polymerases and numerous methods of using the subject DNA polymerases, including uses for chain termination sequencing and PCR. The present teachings relate to DNA polymerases that have an F667Y mutation as defined with respect to Taq DNA polymerase and exhibit reduced discrimination against labeled nucleotides into polynucleotides. The teachings also include kits comprising the subject DNA polymerases and numerous methods of using the subject DNA polymerases, including uses for chain termination sequencing and PCR.

DNA polymerases having improved labeled nucleotide incorporation properties

An improved landing craft includes a hull that supports bow and stern ramps for enabling vehicles to drive through during loading and unloading. The drive-through arrangement also enables the crew to load and unload the craft much faster and more easily than existing vessels. A ballasting arrangement can be used to trim the hull when it is to be beached or to be extracted and/or during loading/unloading.

Planing landing craft

The present invention relate to methods and compositions for simultaneously analyzing multiple different polynucleotides of a polynucleotide composition comprising multiple diverse polynucleotide sequences. The subject methods and compositions may also be applied to analyze or identify single polynucleotides; however, the subject methods and compositions are particularly useful for analyzing large diverse populations of polynucleotides, e.g., cDNA libraries. Most embodiments of the invention involve hybridizing terminus probes (of known base sequence) to adapter-modified restriction fragment generated from polynucleotide for analysis, and subsequently joining the terminus probes and internal fragment probes to each other. The terminus probe hybridizes to bases of restriction endonuclease recognition site present at the terminus of a restriction fragment generated from the polynucleotide for analysis. The terminus probes and internal fragment probes may be marked so as to facilitate the simultaneous testing of multiple polynucleotides for the presence of many possible nucleotide base sequences. The identity or expression of a particular polynucleotide of interest may be ascertained (or at least partially determined) by producing a short identifier sequence derived from the nucleotide base sequence information obtained from (1) the hybridization of a terminus probe, and (2) the recognition site of a restriction endonuclease used to generate the polynucleotide molecule of interest. Multiple identification sequences may be obtained in parallel, thereby permitting the rapid characterization of a large number of diverse polynucleotides. Parallel processing may be achieved by differentially marking terminus probes or internal fragment probes. Parallel processing may be achieved by using ordered arrays of oligonucleotides that are terminus probes.

Restriction enzyme mediated adapter

The present invention relate to methods and compositions for simultaneously analyzing multiple different polynucleotides of a polynucleotide composition comprising multiple diverse polynucleotide sequences. The subject methods and compositions may also be applied to analyze or identify single polynucleotides; however, the subject methods and compositions are particularly useful for analyzing large diverse populations of polynucleotides, e.g., cDNA libraries. Most embodiments of the invention involve hybridizing terminus probes (of known base sequence) and internal fragment probes (of known base sequence) at adjacent positions on an adapter-modified restriction fragment generated from polynucleotide for analysis, and subsequently joining the terminus probes and internal fragment probes to each other. The terminus probe hybridizes to bases of restriction endonuclease recognition site present at the terminus of a restriction fragment generated from the polynucleotide for analysis. Internal fragment probes hybridizes to the same strand of the restriction fragment that the terminus probe hybridizes to and hybridizes to the restriction fragment portion of adapter-modified representative restriction fragments. The terminus probes and internal fragment probes may be marked so as to facilitate the simultaneous testing of multiple polynucleotides for the presence of many possible nucleotide base sequences. The identity or expression of a particular polynucleotide of interest may be ascertained (or at least partially determined) by producing a short identifier sequence derived from the nucleotide base sequence information obtained from (1) the hybridization of a terminus probe and an internal fragment probe, each of known base sequence, at adjacent positions on a polynucleotide of interest, and (2) the recognition site of a restriction endonuclease used to generate the polynucleotide molecule of interest. Multiple identification sequences may be obtained in parallel, thereby permitting the rapid characterization of a large number of diverse polynucleotides. Parallel processing may be achieved by differentially marking terminus probes or internal fragment probes. Parallel processing may be achieved by using ordered arrays of oligonucleotides that are terminus probes.

Adapter directed expression analysis

The method of synthesizing a segment of DNA, typically a gene, of predetermined structure which comprises synthesizing a first pair of oligonucleotides having ends complementary to the ends enzymatically made at a restriction site on a standard plasmid vector, said first pair of oligonucleotides including adjacent their ends portions of said predetermined structure and each having a pair of restriction sites internal to said portions of predetermined structure which restriction sites are unique to the enzyme and not present in the plasmid vector, cloning said first pair of oligonucleotides into said standard plasmid vector, and amplifying said plasmid vector in vivo to recover plasmid vector containing said first pair of oligonucleotides, repeating said method with a second pair of ologonucleotides, containing portions of predetermined structure which are adjacent to the portions present in the first pair of oligonucleotides, and continuing said repetition until all of said segment of predetermined structure has been cloned into said standard vector and amplified.

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Bradbury, CA, United States of America

John H Richards