Patent ID: 6136568
Filing Date: 2000-10-24
Classification: C12N,Y10S

Abstract:
A method for synthesizing a polynucleotide molecule having a defined sequence, comprising repeatedly performing a cycle of steps:(a) annealing a primer having 5' and 3' ends with a template having a 5' region non-complementary to t he primer, a 3' region complementary to the 3' end of the primer, and a non-reactive 3' terminus, the template being shorter than the primer;(b) reacting the primer with at least one nucleotide in the presence of a template-dependent polynucleotide polymerase to produce an extended primer including at its 3' end at least one nucleotide complementary to the 5' region of the template; and(c) dissociating the template from the extended primer;wherein for each repetition of the cycle the extended primer of step (c) is used as the primer for a next repetition of step (a), the template may be the same as or different from a template of a previous repetition, and the template dependent polynucleotide polymerase may be the same as or different from a template dependent polynucleotide polymerase of a previous repetition.