METHOD FOR CATTLE GENOTYPING BY BETA-LACTOGLOBULIN A AND B ALLELES (RS109625649 POLYMORPHISM) BASED ON PCR WITH ALLELE-SPECIFIC PROBES Russian patent published in 2022 - IPC C12N1/00 C12Q1/68 

FIELD: biotechnology.

SUBSTANCE: invention relates to the field of biotechnology, molecular genetic diagnostics, genetics and breeding of farm animals, in particular to the assessment of allelic polymorphism of beta-lactoglobulin of cattle by molecular genetic research method. In the method for cattle genotyping by beta-lactoglobulin A and B alleles (rs109625649 polymorphism) based on PCR with allele-specific probes, the following are used:

direct primer F: 5´-GCATGGAGAACAGTGCTGAGC-3´

reverse primer R: 5´-GCAGCGAGGACCACACAG-3´

BLGB - allele-specific fluorescent-labeled probe

5´-FAM- TGGC AGGCC AGGC-BHQ1-3

BLG_A - allele-specific fluorescent-labeled probe

5´-VIC- TGGC AGACCAGGCT-BHQ1-3´.

For homozygous samples by allele B, an increase in fluorescence along the FAM channel is detected. For homozygous samples by allele A, a signal is detected via the VIC channel. For a heterozygous sample (AB genotype), an increase in fluorescence is observed along both detection channels. The presence of two dyes FAM and VIC makes it possible to uniquely determine the presence of each of the studied βLG alleles in the analyzed DNA sample and, accordingly, the genotype of the animal.

EFFECT: invention makes it possible to expand the arsenal of methods for genotyping cattle by alleles A and B βLG (polymorphism rs109625649) based on real-time PCR with allele-specific probes of the TaqMan type, as well as simplify the analysis and reduce its time to 1 hour.

1 cl, 2 dwg