METHOD FOR SAMPLE PREPARATION OF SARS-CoV-2 CORONAVIRUS ISOLATES AND OLIGONUCLEOTIDE PRIMERS FOR ITS IMPLEMENTATION Russian patent published in 2021 - IPC C12N15/00 G01N33/53 

FIELD: biotechnology, medicine, molecular biology, genetic engineering and virology.

SUBSTANCE: invention relates to biotechnology, medicine, molecular biology, genetic engineering and virology. A method for sample preparation of SARS-CoV-2 coronavirus isolates is described, including the stages: (a) reverse transcription of the RNA of the virus; (b) PCR using two pools of oligonucleotide primers, where oligonucleotide primers having the structure SEQ ID NO: 1-4 are used for the first pool, with a concentration of each primer in the reaction mixture of 0.1 pmol/mcl, and for the second pool, oligonucleotide primers are used having the structure of SEQ ID NO: 5-10, with a concentration in the reaction mixture of each primer of 0.2 pmol/mcl; (c) evaluating the PCR products obtained during the amplification by electrophoresis; (d) combining PCR products of the two pools; (e) purifying amplicons from the reaction mixture using paramagnetic particles with a polymer carboxylated coating; (f) indexing using oligonucleotides containing sequences complementary to the adapter sequences SEQ ID NO: 11-12, and index sequences; (g) re-cleaning from the reaction mixture using paramagnetic particles with a polymer carboxylated coating. Provides a pool of oligonucleotide primers for implementing the method according to claim 1, having the structure of SEQ ID NO: 1-4, for performing the function of primers complementary to the regions of the gene encoding the SARS-CoV-2 S protein, and a pool of oligonucleotide primers for implementing the method according to claim 1, having the structure of SEQ ID NO: 5-10, to act as primers complementary to the regions of the gene encoding the SARS-CoV-2 S protein.

EFFECT: invention makes it possible to carry out sample preparation of libraries containing target fragments of the SARS-CoV-2 genome containing known epidemiologically significant mutations, with the least labor, time and material costs, without reducing the quality and volume of the studies performed due to the use.

9 cl, 2 dwg, 1 tbl, 3 ex