Compositions for enzyme catalyzed template-independent formation of phosphodiester bonds using protected nucleotides
First Claim
1. A composition, comprising:
- (a) a catalytic amount of a template independent enzyme; and
(b) nucleoside 5′
-triphosphate having a removable blocking moiety protecting the 3′
position of said nucleoside 5′
-triphosphate.
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Accused Products
Abstract
A method for the stepwise creation of phosphodiester bonds between desired nucleosides resulting in the synthesis of polynucleotides having a predetermined nucleotide sequence by preparing an initiation substrate containing a free and unmodified 3′-hydroxyl group; attaching a mononucleotide selected according to the order of the predetermined nucleotide sequence to the 3′-hydroxyl of the initiating substrate in a solution containing a catalytic amount of an enzyme capable of catalyzing the 5′ to 3′ phosphodiester linkage of the 5′-phosphate of the mononucleotide to the 3′-hydroxyl of the initiating substrate, wherein the mononucleotide contains a protected 3′-hydroxyl group, whereby the protected mononucleotide is covalently linked to the initiating substrate and further additions are hindered by the 3′-hydroxyl protecting group. Methods in which a mononucleotide immobilized on a solid support is added to a free polynucleotide chain are also disclosed.
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Citations
7 Claims
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1. A composition, comprising:
-
(a) a catalytic amount of a template independent enzyme; and
(b) nucleoside 5′
-triphosphate having a removable blocking moiety protecting the 3′
position of said nucleoside 5′
-triphosphate.- View Dependent Claims (2, 3, 4, 5, 6, 7)
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Specification