3' protected nucleotides for enzyme catalyzed template-independe

Details 3' protected nucleotides for enzyme catalyzed template-independe 3' protected nucleotides for enzyme catalyzed template-independe

1995-06-07

1998-06-09

Leary, Louise N.

Organic compounds -- part of the class 532-570 series

Organic compounds

Carbohydrates or derivatives

536 251, 536 2531, 536 2532, 536 2533, 536 2534, 536 261, 435 6, C07H 2100, C07H 2102, C07H 1904, C12Q 168

Patent

active

057635943

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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