Recovery of oligonucleotides by gas phase cleavage
First Claim
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1. A method for recovering oligonucleotides from a solid support wherein said oligonucleotides are the synthetic products of cyanoethyl phosphoramidite oligonucleotide synthesis comprising the steps of:
- (a) providing a solid support having synthesized oligonucleotides attached thereto, and(b) incubating the solid support in a sealed chamber containing gaseous cleavage/deprotection reagent selected from the group consisting of organic and inorganic amines, under conditions suitable to cleave or deprotect the synthesized oligonucleotides within 90 minutes.
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Abstract
A process for the cleavage and deprotection of newly synthesized oligonucleotides from solid supports, which involves incubating the solid support in an environment comprising gaseous cleavage/deprotection reagent such as gaseous ammonia or ammonium hydroxide vapors. The method lends itself well to the use of supports such as microtiter plates, that can be used to simultaneously perform up to 96 individual synthetic processes.
130 Citations
19 Claims
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1. A method for recovering oligonucleotides from a solid support wherein said oligonucleotides are the synthetic products of cyanoethyl phosphoramidite oligonucleotide synthesis comprising the steps of:
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(a) providing a solid support having synthesized oligonucleotides attached thereto, and (b) incubating the solid support in a sealed chamber containing gaseous cleavage/deprotection reagent selected from the group consisting of organic and inorganic amines, under conditions suitable to cleave or deprotect the synthesized oligonucleotides within 90 minutes. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12)
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13. A method for recovering synthesized oligonucleotides from a solid support, the method comprising the steps of:
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(a) providing a solid support selected from the group consisting of solid, porous or hollow bead particles, permeable membranes, impermeable membranes, and stable emulsified droplets, the support having synthesized oligonucleotide attached thereto, (b) incubating the solid support in a sealed chamber containing gaseous cleavage/deprotection reagent selected from the group consisting of ammonia, ammonium hydroxide and methyl amine under conditions suitable to cleave or aleprotect the synthesized oligonucleotide within 90 minutes. - View Dependent Claims (14, 15, 16, 17, 18)
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19. A method for recovering synthesized oligonucleotides from a solid support, the method comprising the steps of:
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(a) providing a solid support selected from the group consisting of controlled pore glass and polystyrene, the support having the synthetic oligonucleotide product of a cyanoethyl phosphoramidite process attached thereto, (b) incubating the solid support in a sealed chamber containing gaseous cleavage/deprotection reagent selected from the group consisting of ammonia, ammonium hydroxide and methyl amine under conditions suitable to cleave or deprotect the synthesized oligonucleotide within 90 minutes, and (c) resuspending the cleaved/deprotected oligonucleotide in aqueous solution.
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Specification