Process for detecting or quantifying nucleic acids in a library

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 comprising the steps of:
a) providing:
(i) a first 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;(iii) polymerizing means for synthesizing nucleic acid copies of said nucleic acid analytes, said polymerizing means comprising a) a second array comprising a first set of primers, wherein said first set of primers are complementary, in part or in whole, to sequences of said nucleic acids of interest, and b) a second set of primers, wherein said first set of primers are fixed or immobilized to the second array, and wherein said second set of primers comprises at least two segments, the first segment at the 3′ end comprising random sequences, and the second segment comprising at least one RNA promoter; and(iv) means for synthesizing labeled RNA copies from said RNA promoter; b) contacting said library of nucleic acid analytes with said first set of primers on the second array to form more than one first bound entity; c) extending said bound first set of primers on the second array by means of template sequences provided by said nucleic acid analytes to form first copies of said analytes; c′) separating or removing said template sequences from said first copies; d) contacting said first copies on the second array 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 first copies to form more than one complex comprising first copies and second copies formed by extension of said second set of primers; f) synthesizing labeled RNA copies from the RNA promoter in said second copies; g) hybridizing said RNA copies formed in step f) to said first array of nucleic acids provided in step a) (i); and h) detecting or quantifying any of said copies hybridized to said first array in step g).