Analyzing nucleic acid mixture by hybridization to an array, useful e.g. for expression analysis, using sample mixture of labeled restriction fragments of uniform size

摘要

Method for analyzing a nucleic acid (NA) mixture by hybridization to a DNA array of immobilized NA, which uses a labeled NA mixture of reduced complexity as the hybridization sample. Method for analyzing a nucleic acid (NA) mixture by hybridization to a DNA array of immobilized NA, which uses a labeled NA mixture of reduced complexity as the hybridization sample. The method comprises: (a) cleaving an NA mixture with at least one BcgI-like restriction endonuclease (RE); (b) labeling the fragments, of (essentially) the same lengths; (c) optionally isolating the mixture of fragments; (d) labeling the fragments; (e) optionally isolating the labeled fragments; (f) hybridizing the mixture of labeled fragments to a DNA array; and (g) evaluating hybridization. Independent claims are also included for the following: (1) several variants of the method; (2) mixture of labeled NA fragments of (essentially) the same length prepared by the new method; and (3) DNA array comprising immobilized oligonucleotides (ON) of essentially uniform length (20-100 nucleotides) and having sequences identical with those of fragments obtained by digestion with at least one RE.