Process for the preparation of oligo- and polydeoxyribonucleotides
AssigneesVepex Contractor Ltd., Mta Szegedi Biologiai Kozpontja
InventorsAndras Simoncsits, Miklos Kalman, Csaba Kari, Imre Cserpan
Life of this patent
Estimating the term…
Abstract
The invention relates to a process for the preparation of oligo- and polydeoxyribonucleotides by synthesizing the complementary strand of a single-stranded DNA piece enzymatically, in the presence of deoxyribonucleoside 5'-triphosphates in a cloning vector.
Claims10
A process for the preparation of oligo- and polydeoxyribonucleotides comprising inserting a single-stranded DNA piece into a cloning vector, said vector containing a number of closely located recognition sequences of restriction enzymes giving a 3'-protruding end, each present only once in the said vector and which, following cleavage with two corresponding restriction enzymes, produces a linearized vector containing a 5'-protruding end and a 3'-protruding end, said single-stranded DNA piece being inserted into the linearized vector such that, after ligation, the linear vector containing said single-stranded DNA contains 5'-protruding regions at both ends, and synthesizing the complementary strand of said single-stranded DNA piece entirely enzymatically in the presence of deoxyribonucleoside 5'-triphosphates in said cloning vector.
An adapter having a double-stranded region, a 5'-protruding end complementary to a unique cleavage site of the vector which results in a 5'-protruding end, and a 3'-protruding end characteristic of a restriction enzyme the recognition sequence of which is not present in the linearized vector, and from which sequence at most one or two nucleotides remain in the resulting DNA region at the site of attachment of the cloned DNA fragments following enzymatic synthesis and cloning.
Dependencies are inferred from the claim text.