FIELD: biotechnology.
SUBSTANCE: invention relates to the field of biotechnology, specifically to means for determining the CCR5delta32 mutation in the human genome, and can be used to determine and count the number of mutant CCR5delta32 alleles in the studied genetic material during genetic screening and to determine the efficiency of genome editing when using innovative methods of treating HIV infections. A kit has been proposed for determining the CCR5delta32 mutation in the human genome during digital drop multiplex PCR in real time, including forward and reverse primers, a CCR5-78-MT-FAM fluorescent probe with a fluorescent dye molecule 5 (6) -carboxyfluorescein (FAM) covalently stitched to the 5'-end of the sequence and a fluorescence quencher molecule BHQ1 covalently stitched to the 3'-end of the sequence and a CCR5-78-WT-R6G fluorescent probe with covalently 5'-the end of the sequence with a molecule of a fluorescent dye Rhodamine 6G (R6G) and a molecule of a fluorescence quencher BHQ2 covalently stitched to the 3'-end of the sequence.
EFFECT: high-precision counting of CCR5-Δ32 copy number of mutant alleles with a resolution of up to 0.1% due to the high specificity of the reagents of the proposed set.
1 cl, 4 dwg, 1 tbl
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Authors
Dates
2021-06-02—Published
2020-10-27—Filed