Hybridization compositions and methods

Abstract

The invention provides methods and compositions for hybridizing at least one molecule to a target. The invention may, for example, utilize a of cyclic and/or non-cyclic solvent that is non-toxic and may eliminate or reduce the amount of formamide in the hybridization composition.

1 - 57 . (canceled) 
     
     
         58 . A method of hybridizing nucleic acid sequences comprising:
 providing a first nucleic acid sequence,   providing a second nucleic acid sequence,   providing a hybridization composition comprising at least one solvent in an amount effective to denature double-stranded nucleotide sequences, and   combining the first and the second nucleic acid sequence and the hybridization composition for at least a time period sufficient to hybridize the first and second nucleic acid sequences,   wherein the solvent is chosen from butadiene sulfone, tetrahydrothiophene I-oxide (tetramethylene sulfoxide), and 2-pyrrolidone.   
     
     
         59 . A method of hybridizing nucleic acid sequences comprising:
 providing a first nucleic acid sequence in an in situ biological sample, and   applying a hybridization composition comprising a second nucleic acid sequence and at least one solvent in an amount effective to denature double-stranded nucleotide sequences to said first nucleic acid sequence for at least a time period sufficient to hybridize the first and second nucleic acid sequences,   wherein the solvent is chosen from butadiene sulfone, tetrahydrothiophene I-oxide (tetramethylene sulfoxide), and 2-pyrrolidone.   
     
     
         60 . The method according to  claim 58 , wherein a sufficient amount of energy to hybridize the first and second nucleic acids is provided. 
     
     
         61 . The method according to  claim 60 , wherein the energy is provided by heating the hybridization composition and nucleic acid sequence. 
     
     
         62 . The method according to  claim 60 , wherein the heating is performed by the use of microwaves, hot baths, hot plates, heat wire, peltier element, induction heating or heat lamps. 
     
     
         63 . The method according to  claim 58 , wherein the first nucleic acid sequence is double stranded and the second nucleic acid is single stranded. 
     
     
         64 . The method according to  claim 58 , wherein the denaturation and hybridization occur separately. 
     
     
         65 . The method according to  claim 58 , wherein the hybridizing includes steps of heating and cooling the hybridization composition and nucleic acid sequences. 
     
     
         66 . The method according to  claim 58 , wherein the hybridization takes less than 2 hours. 
     
     
         67 . The method according to  claim 66 , wherein the hybridization takes less than 1 hour. 
     
     
         68 . The method according to  claim 67 , wherein the hybridization takes less than 30 minutes. 
     
     
         69 . The method according to  claim 68 , wherein the hybridization takes less than 15 minutes. 
     
     
         70 . The method according to  claim 69 , wherein the hybridization takes less than 5 minutes. 
     
     
         71 . The method according to  claim 58 , wherein the cooling takes less than 1 hour. 
     
     
         72 . The method according to  claim 71 , wherein the cooling takes less than 30 minutes. 
     
     
         73 . The method according to  claim 72 , wherein the cooling takes less than 15 minutes. 
     
     
         74 . The method according to  claim 73 , wherein the cooling takes less than 5 minutes. 
     
     
         75 . The method according to  claim 58 , wherein the denaturation is performed at 67° C. 
     
     
         76 . The method according to  claim 58 , wherein the first nucleic acid sequence is in a biological sample. 
     
     
         77 . The method according to  claim 76 , wherein the biological sample is a cytology or histology sample.