Patent ID: 6060240
Filing Date: 2000-05-09
Classification: C12N,C12Q

Abstract:
A method of normalizing a genetic library, comprising the steps of:(a) attaching unique oligonucleotide identifier tags to cDNA inserts derived from a genetic library;(b) hybridizing the cDNA inserts of step (a) with cDNA derived from a source of interest under conditions that promote the formation of perfectly matched duplexes, wherein the cDNA is labeled with a first label;(c) contacting the mixture of step (b) with solid supports having attached thereto the complements of the oligonucleotide identifier tags under conditions that promote the formation of perfectly matched duplexes between the oligonucleotide identifier tags and their respective complements in the presence of free oligonucleotide identifier tags labeled with a second label and corresponding in sequence to the oligonucleotide identifier taos of step (a);(d) sorting solid supports according to the relative amount of said first label and said second label, wherein said relative amount of said first and second label correlates with the abundance of a cDNA, or mRNA sequence; and(e) amplifying cDNA inserts present at lower abundance in order to match the abundance of all cDNA sequences such that they are represented at substantially similar levels in the library.