Lifespan Biosciences, Inc.

Glenna C Burmer

Joseph P Brown

Amanda A Ford

Christine C Stewart

This invention relates to the discovery of the association of certain nucleic acid sequences and proteins with breast cancer, the use of such sequences as a diagnostic indicator and treatments based on the association.

Breast cancer associated nucleic acid sequences and their associated proteins

This invention provides methods for the isolation of complete representation of cellular messenger ribonucleic acid from wax-embedded tissue samples. Expression of genes of interest can thus be fortuitously determined and cDNA probes can be readily developed.

Methods of measuring gene expression using wax-embedded tissue specimens

The present invention relates generally to high throughput screening methods. More particularly, the present invention provides screening methods that can readily be used to identify simultaneously multiple proteins or compounds that interact with multiple ligands, using a tagged array of ligands.

Tagged ligand arrays for identifying target-ligand interactions

The present invention provides gene subtraction methods that can be used to identify and isolate from hybridization mixtures differentially expressed nucleic acid and/or commonly expressed nucleic acid between two populations of nucleic acid. The methods of the present invention employ a restriction endonuclease recognition site that is also recognized in whole or in part by a second restriction endonuclease (e.g., HinPI and BssHII or HhaI and HinP1I) to create different ends (e.g., sticky ends) between the first and second nucleic acid populations. In addition, the methods of the present invention use a selective ligation step, a biotinylated nucleotide extension step, or both to differentiate between homoduplexes, which allow for the identification of sequences that are different between the two nucleic acid populations, and heteroduplexes, which allow for the identification of sequences that are common to the two nucleic acid populations.

Subtractive hybridization techniques for identifying differentially

The invention disclosed here allows for the differential isolation of nucleic acid sequences that are present in one nucleic acid population and not in another. The method is based upon using restriction endonucleases to digest two populations of nucleic acid, preferably cDNAs, preferably ligating different sets of adaptors to each of the two nucleic acid populations, followed by hybridization, restriction digestion and isolation of the desired molecules. The unique aspects of this invention include the use of a restriction enzyme to isolate the target duplex DNA molecule from a hybridization mixture. Certain embodiments of the invention include the direct or indirect incorporation of a capture molecule or ligand (e.g., biotin, dioxigenin) within the amplified nucleic acid fragments, which allows for a system in which molecules can be rescued from both the captured population as well as the effluent or otherwise uncaptured population.

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Seattle, WA, United States of America

Glenna C Burmer