System for detecting polynucleotides

Abstract

The present invention relates to methods for detecting the presence or amount of a target polynucleotide. A polynucleotide, target nucleic acid analog, and dye are combined to form a mixture. The optical property of the dye is observed after the mixture is exposed to a stimulating means. Optionally, after the stimulating means is employed, the mixture is compared to a reference value characteristic of the rate of change in the optical property of the dye in a similar mixture containing a known amount of a target polynucleotide/nucleic acid analog hybrid to determine a relative rate of change in the optical property. The change in a property of the mixture after exposure thereof to a stimulating means or the relative rate of change in the optical property of dye in the mixture is correlated with the presence or amount of the specified target polynucleotide in the sample.

1 .- 16 . (canceled) 
     
     
         17 . A kit comprising:
 (a) a low pH dye reagent comprising a carbocyanine dye; and   (b) a non-ionic detergent.   
     
     
         18 .- 20 . (canceled) 
     
     
         21 . The kit of  claim 17 , wherein the carbocyanine dye is 3,3′-diethylthiacarbocyanine. 
     
     
         22 . The kit of  claim 17 , further comprising a nucleic acid analog stock solution. 
     
     
         23 . The kit of  claim 23 , wherein the nucleic acid analog stock solution is a peptide nucleic acid (PNA) stock solution. 
     
     
         24 . The kit of  claim 17 , further comprising a vehicle to facilitate hybridization of a nucleic acid analog to a target polynucleotide. 
     
     
         25 . The kit of  claim 17 , wherein the non-ionic detergent is Tween 80. 
     
     
         26 . The kit of  claim 17 , further comprising an anti-microbial agent. 
     
     
         27 . The kit of  claim 17 , further comprising a diluting buffer. 
     
     
         28 . The kit of  claim 17 , further comprising a source of light stimulus. 
     
     
         29 . The kit of  claim 17 , further comprising at least one polynucleotide manipulating component. 
     
     
         30 . The kit of  claim 17 , further comprising at least one kit component selected from the group consisting of chemical-resistant disposal bags, tubes, diluents, gloves, scissors, marking pens, and eye protection. 
     
     
         31 . A kit comprising:
 (a) a low pH dye reagent comprising a carbocyanine dye, an antimicrobial agent, and Tween 80;   (b) a diluting solution comprising EDTA;   (c) a nucleic acid analog stock solution comprising PNA and EDTA; and   (d) control DNA.   
     
     
         32 . The kit of  claim 31 , wherein the carbocyanine dye is 3,3′-diethylthiacarbocyanine.