METHOD TO GENERATE OR DETERMINE NUCLEIC ACID TAGS CORRESPONDING TO THE TERMINAL ENDS OF DNA MOLECULES USING SEQUENCES ANALYSIS OF GENE EXPRESSION (TERMINAL SAGE)

摘要

We describe a method of providing an indication of an instance of expression of a gene, the method comprising the steps of (a) providing a complementary deoxyribonucleic acid (cDNA) having a terminus comprising a terminal transcribed sequence of a gene; (b) linking the cDNA to an linker sequence thereby forming a linked nucleic acid, in which the linker sequence comprises a first recognition site for a first nucleic acid cleavage enzyme, preferably a restriction endonuclease, that allows nucleic acid cleavage at a site distant from the first recognition site; and (c) cleaving the linked nucleic acid with the first nucleic acid cleavage enzyme to provide a linked tag, in which the linked tag comprises a nucleotide sequence tag representative of a terminal transcribed sequence of the gene; and (d) detecting the presence or identity of the linked tag or the nucleotide sequence tag to provide an indication of an instance of gene expression.