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 in the presence of one or more carbohydrates and/or sugars 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 1 , wherein said nucleophilic amino compound is ammonia.
4. The process of claim 3 , wherein said ammonia is present at a psi of about 20 to 200.
5. The process of claim 3 , wherein said ammonia is present at a psi of about 80.
6. The process of claim 1 , wherein said nucleophilic amino compound is ammonia vapors.
7. The process of claim 1 , wherein said nucleophilic amino compound is a C 1-6 alkylamine.
8. The process of claim 1 , further comprising dissolving the substantially deblocked oligonucleotide in a buffer.
9. The process of claim 1 , wherein said conditions comprise carrying the process at about room temperature to about 150° C.
10. The process of claim 1 , wherein said conditions comprise carrying out the process at about 95° C.
11. The process of claim 1 , wherein said conditions comprise carrying out the process for about 1 min to about 2 hrs.
12. The process of claim 1 , wherein said substantially blocked, detectably labeled oligonucleotide is immobilized on a solid phase.
13. The process of claim 12 , wherein said substantially deblocked oligonucleotide is cleaved from said solid phase under said conditions.
14. 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.
15. The process of claim 14 , wherein said blocked, detectably labeled oligonucleotide is immobilized on a solid phase.
16. The process of claim 15 , wherein said substantially deblocked oligonucleotide is released cleaved from said solid phase.
17. A composition comprising a blocked, detectably labeled oligonucleotide and an effective amount of a nucleophilic amino compound sufficient to deblock substantially said oligonucleotide and one or more carbohydrates and/or sugars.
18. The composition of claim 17 , wherein said detectable label is a fluorescent label.
19. The composition of claim 17 , wherein said nucleophilic amino compound is ammonia.
20. The composition of claim 17 , wherein said nucleophilic amino compound is ammonia vapors.