METHOD FOR CREATING A CIRCULAR FORM OF AN NGS LIBRARY FOR HIGH-THROUGHPUT SEQUENCING ON THE BASIS OF DNBSEQ TECHNOLOGY PLATFORMS Russian patent published in 2021 - IPC C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology. The invention is a method of creating a circular form of an NGS library for high-throughput sequencing on platforms of DNBSEQ technology, including the following steps: mixing an NGS library and a splint oligonucleotide; ligation of the ends of DNA fragments with DNA ligase enzyme; degradation of non-circular DNA by exonuclease enzymes; purification of the obtained circular DNA fragments. In this case, mixing the NGS library and the splint oligonucleotide is carried out in a solution containing NaCl at a concentration of 10 mM. Before the stage of ligation of the ends of DNA fragments, the reaction mixture is heated according to the following program: at a temperature of 95 °С for 2 minutes, at a temperature of 65 °С for 2 minutes, at a temperature of 40 °С for 2 minutes and at a temperature of 4 °С for at least 1 minute. The duration of the stage of ligation of the ends of DNA fragments is at least 1 hour at a temperature of 37 °С.

EFFECT: invention ensures a reduction in the required amount of DNA of the NGS library for its preparation for sequencing on DNBSEQ platforms by using optimized chemical composition and original temperature control programs. The invention improves the quality of the NGS library and the obtained sequencing data by reducing the number of PCR cycles during the creation of the NGS library.

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