Patent ID: 6136537
Filing Date: 2000-10-24
Classification: C12N,C12P,C12Q

Abstract:
A method of analyzing gene expression in a cell or tissue, the method comprising the steps of(a) forming a population of cDNA molecules from mRNA of a cell or tissue;(b) digesting the population of cDNA molecules with at least one restriction endonuclease to produce a population of polynucleotides having predetermined ends;(c) enzymatically removing a segment of nucleotides from each predetermined end of each polynucleotide and ligating the segments from each end together to form a pair of sequence stages for each polynucleotide, wherein said segments are formed by inserting each of said polynucleotides into a cloning site of a vector, the cloning site being flanked by a first type IIs restriction site and a second type IIs restriction site such that a type IIs restriction endonuclease recognizing either said first or second sites cleaves the vector within to said polynucleotide, the first IIs restriction site and the second type IIs restriction site being the same or different and each of the first and second type IIs restriction sites being unique to the vector;(d) determining the nucleotide sequences of a sample of pairs of sequence tags; and(e) tabulating the nucleotide sequences of the pairs of sequence tags to form a frequency distribution of gene expression in the cell or tissue.