METHOD FOR OBTAINING COMBINED DNA FRAGMENTS IN PCR (VARIANTS) Russian patent published in 2012 - IPC C12Q1/68 C12Q1/00 

FIELD: biotechnologies.

SUBSTANCE: the invention is referred to the field of biotechnology. Performance of two PCR rounds is enough for obtaining of the end product. In the first round primary DNA fragments are obtained using multiplexed PCR, in the second round cohesive ends appear in the obtained DNA fragments, fragments fuse and the resulting product is amplified. Aggregate of high-molecular weight DNA or human genomic DNA can be used as primary material in the first round, PCR products from the first round are used in the second round. The technique allows for obtaining of rather precise fragments consisting of exons alone. If there is no necessity for obtaining of compound fragments without inserts, it is proposed to get compound fragments from the aggregate of high-molecular weight DNA in one PCR reaction. Multiplexed DNA using special primers that contain ends that are unique for genomic DNA 5' and that are mutually complementary, by which formation of cohesive ends is achieved in synthesized primary fragments. In this process a pair of primers flanking the common compound fragment, which are fully complementary with external edges of desired compound DNA fragment and intended for its amplification, is used, as well.

EFFECT: fusion of PCR fragments that do not require intermediate treatment, preliminary cloning of primary DNA fragments, and also separate amplification of primary fragments.

2 cl, 7 dwg, 3 ex