METHOD FOR APAF1 POLYMORPHISM DETERMINATION, ASSOCIATED WITH HOLSTEIN CATTLE HH1 FERTILITY HAPLOTYPE Russian patent published in 2017 - IPC C12Q1/00 

FIELD: biotechnology.

SUBSTANCE: method for polymorphism determination for genes associated with cattle fertility, namely with apoptotic protease-activating factor 1 (APAF1) gene, associated with Holstein cattle HH1 fertility haplotype. The method includes PCR-RFLP determination of CT polymorphism→ at 63150400 position (genome assembly UMD 3.1) of APAF1 gene. At that, amplification of the gene fragment containing a mutation is carried out using two primers. The nucleotide substitution leading to the elimination of non-specific BstC8I endonuclease non-specific restriction site located at a distance of 10 bp "down stream" of the investigated mutation, is introduced into the reverse primer followed by restriction PCR products hydrolysis with BstC8I endonuclease. Shorter fragments resulting from restriction correspond to Q allele, while a longer non-restricted fragment corresponds to X allele. Animal genotypes identification is based on the results of PCR-RFLP products electrophoresis in agarose gel.

EFFECT: method can be used in farm animals genetics to detect polymorphism in the cattle APAF1 gene, associated with HH1 fertility haplotype, in order to use the results in population genetics, cattle breeding and selection.

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