Probe/mobility modifier complexes for multiplex nucleic acid detection

Abstract

Compositions and methods for the analysis of multiple nucleic acid target sequences are disclosed. The compositions comprise a probe comprising a target-specific portion for sequence-specific hybridization to a target nucleic acid sequence, and a tag; and a mobility-modifier comprising a tail and a tag complement for binding to the tag. The associated methods generally comprise the steps of providing a sample potentially containing one or more target nucleic acid sequences; providing one or more probes, each probe comprising a target-specific portion and a tag; providing one or more mobility modifiers, each mobility modifier comprising a tag complement and a tail; contacting the probe(s) and the target nucleic acid sequence(s) under conditions effective for sequence-dependent hybridization of the probe(s) and the target nucleic acid sequence(s); contacting the probe(s) and the mobility-modifier(s) under conditions suitable for selectively binding the probe(s) to the mobility modifier(s), thereby forming one or more a probe/mobility modifier complex(s); and analyzing the probe/mobility modifier complex(s) using a mobility-dependent analysis technique.

1 . A binary composition for detecting one or more target nucleic acid sequences comprising: 
 a probe comprising a target-specific portion for sequence-specific hybridization to a target nucleic acid sequence, and a tag; and    a mobility-modifier comprising a tail and a tag complement for binding to the tag.    
     
     
         2 . The composition of  claim 1  wherein the target-specific portion comprises polynucleotide.  
     
     
         3 . The composition of  claim 2  wherein the target-specific portion comprises a 3′-hydroxyl group.  
     
     
         4 . The composition of  claim 2  wherein the target-specific portion comprises PNA.  
     
     
         5 . The composition of  claim 1  wherein the tag portion comprises polynucleotide.  
     
     
         6 . The composition of  claim 5  wherein the tag portion is PNA.  
     
     
         6 . 1  (canceled)  
     
     
         7 . The composition of  claim 1  wherein both the tag and tag complement are polynucleotide, and one of the tag complement and tag comprises a sequence selected from the group consisting of (CAG) n  and (TCC) n  wherein n is 1 to 10.  
     
     
         8 . The composition of  claim 1  wherein the mobility modifier comprises a tail portion.  
     
     
         9 . The composition of  claim 8  wherein the tail is a polymer.  
     
     
         10 . The composition of  claim 9  wherein the polymer is selected from the group consisting of polyethyleneoxide and polypeptide.  
     
     
         11 . The composition of  claim 1  further comprising a hybridization enhancer.  
     
     
         12 - 25 . (canceled)