Methods and materials for assessing rna expression

Abstract

This document provides methods and materials for assessing RNA expression. For example, methods and materials for detecting the presence, absence, or amount of target nucleic acid (e.g., target RNA or target cDNA produced from target RNA), kits for detecting the presence, absence, or amount of target nucleic acid (e.g., target RNA or target cDNA produced from target RNA), and methods for making such kits are provided.

1 . (canceled) 
     
     
         2 . A kit for determining whether a target RNA is present in a sample, wherein said kit comprises:
 (a) cDNA generated from RNA of said sample,   (b) probe nucleic acid comprising a nucleotide sequence complementary to a sequence of a target cDNA of said target RNA, wherein at least a portion of said target cDNA is capable of hybridizing to at least a portion of said probe nucleic acid to form a double-stranded portion of nucleic acid comprising a restriction endonuclease cut site, wherein said probe nucleic acid comprises a restriction endonuclease, and   (b) reporter nucleic acid comprising a label and a double-stranded portion of nucleic acid comprising a restriction endonuclease cut site of said restriction endonuclease of said probe nucleic acid.   
     
     
         3 . The kit of  claim 2 , wherein said probe nucleic acid is single-stranded probe nucleic acid. 
     
     
         4 . The kit of  claim 2 , wherein said kit comprises a solid support, and wherein said probe nucleic acid is attached to said solid support. 
     
     
         5 . The kit of  claim 4 , wherein a portion of said probe nucleic acid comprising said restriction endonuclease is releasable from said solid support via cleavage with a recognition restriction endonuclease having the ability to cleave at said restriction endonuclease cut site formed by said target cDNA and said portion of said probe nucleic acid. 
     
     
         6 . The kit of  claim 2 , wherein said kit further comprises a recognition restriction endonuclease having the ability to cleave at said restriction endonuclease cut site formed by said target cDNA and said portion of said probe nucleic acid. 
     
     
         7 . The kit of  claim 2 , wherein said probe nucleic acid comprise (i) a single-stranded portion comprising said nucleotide sequence complementary to said sequence of said target cDNA and (ii) a double-stranded portion. 
     
     
         8 . The kit of  claim 7 , wherein said probe nucleic acid comprises a first nucleic acid strand comprising said nucleotide sequence complementary to said sequence of said target cDNA hybridized to a second nucleic acid strand comprising said restriction endonuclease. 
     
     
         9 . The kit of  claim 2 , wherein said probe nucleic acid is lyophilized. 
     
     
         10 . The kit of  claim 2 , wherein said kit comprises a solid support, and wherein said reporter nucleic acid is attached to said solid support. 
     
     
         11 . The kit of  claim 10 , wherein said reporter nucleic acid is directly attached to said solid support. 
     
     
         12 . The kit of  claim 2 , wherein said reporter nucleic acid comprises a single-stranded portion of nucleic acid. 
     
     
         13 . The kit of  claim 2 , wherein said label is a fluorescent label, a radioactive label, an enzyme label, or a redox label. 
     
     
         14 . The kit of  claim 2 , wherein said restriction endonuclease is an EcoRI endonuclease. 
     
     
         15 . The kit of  claim 2 , wherein said restriction endonuclease is a BamHI endonuclease. 
     
     
         16 . The kit of  claim 2 , wherein said kit is a microfluidic device. 
     
     
         17 . The kit of  claim 2 , wherein said kit further comprises a heating mechanism configured to initiate an exothermic chemical reaction within said kit. 
     
     
         18 . The kit of  claim 2 , wherein said kit further comprises a cooling mechanism configured to initiate an endothermic chemical reaction within said kit. 
     
     
         19 . The kit of  claim 2 , wherein said kit further comprises a temperature indicator.