Method for deblocking of labeled oligonucleotides
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 1-6 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. The process of claim 1 , further comprising wherein said substantially blocked, detectably labeled oligonucleotide is immobilized on a solid phase.
14. The process of claim 13 , further comprising wherein said substantially deblocked oligonucleotide is released from said solid phase under said conditions.
15. 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.
16. The process of claim 15 , further comprising wherein said blocked, detectably labeled oligonucleotide is immobilized on a solid phase.
17. The process of claim 16 , further comprising wherein said substantially deblocked oligonucleotide is released from said solid phase.
18. A composition comprising a blocked, detectably labeled oligonucleotide and an effective amount of a nucleophilic amino compound sufficient to deblock substantially said oligonucleotide.
19. The composition of claim 18 , wherein said detectable label is a fluorescent label.
20. The composition of claim 18 , wherein said nucleophilic amino compound is ammonia.