3' Biased Detection of Nucleic Acids
The invention provides materials and methods for the detection of nucleic acid expression via the 3′ portion of expressed sequences. Embodiments of the invention include the use of microarrays comprising nucleic acid probes that are complementary to the 3′ end of expressed sequences and by the use of quantitative PCR (Q-PCR) based amplification of sequences found at or near the 3′ end of expressed sequences. The invention may be used to detect the presence of expressed nucleic acids encoding particular gene products (sequences present in a “transcriptome”).
1 - 19 . (canceled)
20 . A method of analyzing gene expression in a cell, comprising preparing a polynucleotide comprising gene sequences expressed in said cell and hybridizing said polynucleotide to a microarray comprising at least 5 oligonucleotide probes, each of 150 nucleotides or less in length, and complementary to at least 10 consecutive nucleotides of an mRNA molecule, wherein said at least 10 consecutive nucleotides is, in its entirety, less than 360 nucleotides from the site of poly(A) addition of said mRNA molecule.
21 . The method of claim 20 wherein said cell is from an FFPE sample.
22 . The method of claim 20 wherein said cell is a breast cancer cell.
23 . The method of claim 20 wherein said cell has been isolated by microdissection of a cell containing sample.
24 . A method of determining the presence of breast cancer in a subject, comprising preparing a polynucleotide comprising gene sequences expressed in a breast cancer cell of a cell containing sample from said subject, and hybridizing said amplified sequences to a microarray comprising at least 5 oligonucleotide probes, each of 150 nucleotides or less in length, and complementary to at least 10 consecutive nucleotides of an mRNA molecule, wherein said at least 10 consecutive nucleotides is, in its entirety, less than 360 nucleotides from the site of poly(A) addition of said mRNA molecule.
25 . The method of claim 24 wherein said sample is a biopsy.
26 . The method of claim 24 wherein said sample is obtained by fine needle aspiration or ductal lavage.
27 . The method of claim 24 wherein said cell has been isolated by microdissection of said cell containing sample.
28 . The method of claim 23 wherein said sample is a human sample.
29 - 40 . (canceled)