Processes for detecting or quantifying nucleic acids using an array of fixed or immobilized nucleic acids

摘要

This invention provides novel compositions and processes for analyte detection, quantification and amplification. Nucleic acid arrays and libraries of analytes are usefully incorporated into such compositions and processes. Universal detection elements, signaling entities and the like are employed to detect and if necessary or desirable, to quantify analytes. Amplification of target analytes are also provided by the compositions and processes of this invention.

主权项

1. A process for detecting or quantifying more than one nucleic acid of interest in a library, said process comprising the steps of:
a) providing:
(i) an array of fixed or immobilized nucleic acids complementary in part or whole to sequences of said nucleic acids of interest;(ii) a library of nucleic acid analytes which may contain the nucleic acids of interest sought to be detected or quantified; and(iii) polymerizing means for synthesizing nucleic acid copies of said nucleic acid analytes, said polymerizing means comprising a first set of primers and a second set of primers; wherein said first set of primers consists of sequences that are complementary to inherent UDTs, said inherent UDTs consisting of 3′ poly A segments, consensus sequences, or both, wherein the consensus sequence is selected from a sequence for poly A addition, splicing elements, multicopy repeats, or a combination of any of the foregoing, and wherein said second set of primers comprises at least two segments, the first segment at the 3′ end comprising random sequences comprising at least a hexameric sequence, and the second segment comprising at least one production center comprising an RNA promoter;(iv) means for synthesizing nucleic acid copies by transcription; b) contacting said library of nucleic acid analytes with said first set of primers to form more than one first bound entity; c) extending said bound first set of primers by means of template sequences provided by said nucleic acid analytes to form first copies of said analytes; d) contacting said extended first copies with said second set of primers to form more than one second bound entity; e) extending said bound second set of primers by means of template sequences provided by said extended first copies to form more than one complex comprising extended first copies and extended second set of primers; f) transcribing said complex from an RNA promoter in said second set of primers in said complexes; g) hybridizing said nucleic acid copies formed in step f) to said array of nucleic acids provided in step a) (i); and h) detecting or quantifying any of said hybridized copies obtained in step g).