METHOD FOR CREATING THE NGS RING FORM OF THE ILLUMINA LIBRARY FOR HIGH-PERFORMANCE SEQUENCING ON DNBSEQ TECHNOLOGY PLATFORMS Russian patent published in 2022 - IPC C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention relates to biotechnology. A method for creating a ring form of the NGS library for high-performance sequencing on DNBSEQ technology platforms is described, including mixing the NGS library and a splint oligonucleotide; ligation of the ends of DNA fragments by a DNA ligase enzyme; degradation of DNA that is not in ring form by exonuclease enzymes; purification of the obtained ring DNA fragments. At the same time, the NGS library prepared for the Illumina platform is pre-amplified using primers: 5'/5Phos/AATGATACGGCGACCACCGAG-3' and 5'-CAAGCAGAAGACGGCATACGAG-3'. Then it is mixed with splint oligonucleotide 5'-TCGCCGTATCATTCAAGC AGAAGAGG-3'. Before the stage of ligation of the ends of DNA fragments, the reaction mixture is temperature treated according to the following program: at a temperature of 95°C for 2 minutes, at a temperature of 45°C for 2 minutes, at a temperature of 30°C for 2 minutes and at a temperature of 4°C for at least 1 minute.

EFFECT: creating the possibility of sequencing samples existing in the form of Illumina NGS libraries on the DNBSEQ platform; reducing the required number of NGS libraries of the Illumina platform to prepare it for sequencing on DNBSEQ platforms by using original temperature control programs.

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