Methods and compositions for detection and analysis of polynucleotides using light harvesting multichromophores

Abstract

Methods compositions and articles of manufacture for assaying a sample for a target polynucleotide are provided. A sample suspected of containing the target polynucleotide is contacted with a polycationic multichromophore and a sensor polynucleotide complementary to the target polynucleotide. The sensor polynucleotide comprises a signaling chromophore to receive energy from the excited multichromophore and increase emission in the presence of the target polynucleotide. The methods can be used in multiplex form. Kits comprising reagents for performing such methods are also provided.

What is claimed is: 
     
       1. A light harvesting multichromophore system, the system comprising
 a) a signaling chromophore; and   b) a water-soluble conjugated polymer comprising a delocalized electronic structure, wherein the polymer can transfer energy from its excited state to the signaling chromophore to provide a greater than 4 fold increase in fluorescence emission from the signaling chromophore than can be achieved by direct excitation.   
     
     
       2. The light harvesting multichromophore system of  claim 1 , wherein the polymer can transfer energy from its excited state to a signaling chromophore to provide an 8 fold increase in fluorescence emission from the signaling chromophore than can be achieved by direct excitation. 
     
     
       3. The light harvesting multichromophore system of  claim 1 , wherein the signaling chromophore comprises a fluorescent dye. 
     
     
       4. The light harvesting multichromophore system of  claim 1 , wherein the conjugated polymer has the structure 
       
         
           
           
               
               
           
         
       
       
        
       
     
     
       5. The light harvesting multichromophore system of  claim 1 , wherein the system further comprises a substrate. 
     
     
       6. The light harvesting multichromophore system of  claim 1 , wherein the system further comprises a second signaling chromophore. 
     
     
       7. The light harvesting multichromophore system according to  claim 6 , wherein the second signaling chromophore comprises a fluorescent dye. 
     
     
       8. The light harvesting multichromophore system according to  claim 6 , wherein the second signaling chromophore is attached to a substrate. 
     
     
       9. The light harvesting multichromophore system according to  claim 1 , further comprises an excitation source. 
     
     
       10. The light harvesting multichromophore system according to  claim 9 , wherein the excitation source comprises a laser. 
     
     
       11. The light harvesting multichromophore system according to  claim 1 , wherein the system further comprises a detector. 
     
     
       12. The light harvesting multichromophore system according to  claim 11 , wherein the detector comprises a fluorometer. 
     
     
       13. An aqueous composition comprising a light harvesting multichromophore system comprising (a) a signaling chromophore in solution, and (b) a water-soluble conjugated polymer comprising a delocalized electronic structure, wherein the polymer can transfer energy from its excited state to the signaling chromophore to provide a greater than 4 fold increase in fluorescence emission from the signaling chromophore than can be achieved by direct excitation.  
     
     
       14. The aqueous composition of claim 13, wherein the polymer can transfer energy from its excited state to a signaling chromophore to provide an 8 fold increase in fluorescence emission from the signaling chromophore than can be achieved by direct excitation.  
     
     
       15. The aqueous composition of claim 13, wherein the signaling chromophore comprises a fluorescent dye.  
     
     
       16. The aqueous composition of claim 13, wherein the conjugated polymer has the structure 
       
         
           
           
               
               
           
         
       
       wherein n is an integer in the range of 2 to 100,000.  
     
     
       17. The aqueous composition of claim 13, wherein the system further comprises a second signaling chromophore.  
     
     
       18. The aqueous composition of claim 17, wherein the second signaling chromophore comprises a fluorescent dye.