Inventors:
Kenneth Marians - New York NY
Joing Liu - Shanghai, CN
Assignee:
Sloan-Kettering Institute for Cancer Research - New York NY
International Classification:
C12P 1934
US Classification:
435 912, 435 6, 435 911, 536 231, 536 243, 536 2433, 536 253, 536 2532
Abstract:
A method is provided for replicating DNA, and in particular for replicating large segments of DNA. A primer is combined with a target DNA molecule to be replicated. The primer is designed to be at least partially homologous to a known site on the target DNA, and to create a D-loop when hybridized with that site. A replisome is then assembled at the D-loop, and this replisome creates a copy of the DNA, starting at the primer binding site. By utilizing two species of D-loop primers which bind to remote sites on the DNA flanking a region to be replicated, large sections of DNA can be replicated in a manner comparable to PCR. The replicated DNA can be analyzed to detect variations in the genetic sequence of the target, for linkage mapping and as a source of longer DNA molecules having a desired sequence.
