Genotyping of Two Major Genes in Livestock by High Resolution Melt

Document Type : Research Articles

Authors

Tahereh Hosseinzadeh ¹

Fatemeh Bahri Binabaj ²

Ali Javadmanesh ¹

¹ Department of Animal Science, Faculty of Agriculture, Ferdowsi University of Mashhad, Iran

² Department of Animal Science, Khorasan Razavi Agricultural and Natural Resources Research and Education Center, ARREEO, Mashhad, Iran

https://doi.org/10.22067/jcmr.2025.90137.1124

Abstract

Identification of genes affecting production is one of the important priorities in animal breeding strategies. The introgression of genes with a larger effect on quantitative traits into livestock can improve protein and economic resources. The present study aimed to evaluate high-resolution melting (HRM) analysis for genotyping bone morphogenetic protein receptor type 1B (Fecundity Booroola gene or FecB) in sheep and myostatin (MSTN) in cattle. This study was conducted on blood samples from 19 cattle and 170 sheep including: 19 control samples with approved FecB genotypes and 151 samples with unknown genotypes. After DNA extraction from blood and semen samples, a 176-bp fragment of the MSTN gene in cattle and a 140-bp fragment of the FecB gene in sheep were amplified. HRM was optimized and conducted in all samples then, the PCR-RFLP and sequencing methods were done to prove the HRM genotyping results. The results obtained using the HRM method matched those from sequencing and PCR-RFLP, confirming the effectiveness of HRM for genotyping and enabling rapid and accurate identification of individuals with FecB and MSTN mutations. Melting temperature range for MSTN genotypes was between 80.90 and 81.60 ºC and for FecB genotypes was between 79.6 and 80.50 ºC. Utilizing the HRM method for genotyping will provide researchers and commercial animal producers’ access to a low cost, and easier method to genotype for any SNP detection. In this study, we successfully established a genotyping method for two loci in cattle and sheep economically important genes by high-resolution melting method. This method could be used in a large sample size with a high level of confidence.
 

Keywords

High resolution melting

Genotyping

DNA marker

Economic trait

Double muscle

Fecundity

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