Method for deblocking of labeled oligonucleotides

Abstract

The invention relates to a process for deblocking substantially a blocked, detectably labeled oligonucleotide by contacting the blocked detectably labeled oligonucleotide with an effective amount of a nucleophilic amino compound under conditions that result in substantial deblocking of the oligonucleotide, thereby giving the substantially deblocked oligonucleotide.

1 . A process for deblocking substantially a blocked, detectably labeled oligonucleotide comprising contacting the blocked detectably labeled oligonucleotide with an effective amount of a nucleophilic amino compound under conditions that result in the deblocking of the oligonucleotide, thereby giving the substantially deblocked oligonucleotide. 
     
     
         2 . The process of  claim 1 , wherein said detectable label is a fluorescent label. 
     
     
         3 . The process of  claim 2 , wherein said detectable label is hexachlorofluorescein. 
     
     
         4 . The process of  claim 2 , wherein said detectable label is DABCYL. 
     
     
         5 . The process of  claim 1 , wherein said nucleophilic amino compound is ammonia. 
     
     
         6 . The process of  claim 5 , wherein said ammonia is present at a psi of about 20 to 200. 
     
     
         7 . The process of  claim 5 , wherein said ammonia is present at a psi of about 80. 
     
     
         8 . The process of  claim 1 , wherein said nucleophilic amino compound is ammonia vapors. 
     
     
         9 . The process of  claim 1 , wherein said nucleophilic amino compound is a C 16  alkylamine. 
     
     
         10 . The process of  claim 1 , further comprising dissolving the substantially deblocked oligonucleotide in a buffer. 
     
     
         11 . The process of  claim 1 , wherein said conditions comprise carrying the process at about room temperature to about 150° C. 
     
     
         12 . The process of  claim 1 , wherein said conditions comprise carrying the process at about 95° C. 
     
     
         13 - 14 . (canceled) 
     
     
         15 . The process of  claim 1 , wherein said substantially blocked, detectably labeled oligonucleotide is immobilized on a solid phase. 
     
     
         16 . The process of  claim 15 , wherein said substantially deblocked oligonucleotide is released from said solid phase under said conditions. 
     
     
         17 . The process of  claim 16 , wherein said substantially deblocked oligonucleotide is recovered by washing said solid phase with water or a buffer. 
     
     
         18 . A process for deblocking substantially a blocked, detectably labeled oligonucleotide comprising contacting the blocked detectably labeled oligonucleotide with an effective amount of ammonia saturated with water vapor at about 80 psi, 95° C. for about 45 min, thereby giving the substantially deblocked oligonucleotide. 
     
     
         19 - 21 . (canceled) 
     
     
         22 . A composition comprising a blocked, detectably labeled oligonucleotide and an effective amount of a nucleophilic amino compound sufficient to deblock substantially said oligonucleotide. 
     
     
         23 . The composition of  claim 22 , wherein said detectable label is a fluorescent label. 
     
     
         24 . The composition of  claim 22 , wherein said nucleophilic amino compound is ammonia. 
     
     
         25 . The composition of  claim 22 , wherein said nucleophilic amino 15 compound is ammonia vapors.