IN SILICO DESIGN AND VALIDATION OF A NOVEL PCR-RFLP ASSAY FOR GPRC6A RS2247911 POLYMORPHISM INVOLVED IN MALE INFERTILITY | ||||
Bulletin of Pharmaceutical Sciences Assiut University | ||||
Volume 45, Issue 2, December 2022, Page 1077-1084 PDF (1.07 MB) | ||||
Document Type: Original Article | ||||
DOI: 10.21608/bfsa.2022.271821 | ||||
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Authors | ||||
Kenda Jawich ![]() ![]() | ||||
1Department of Biochemistry and Microbiology, Faculty of Pharmacy, Damascus University, Damascus, Syrian Arab Republic | ||||
2Department of Embryology and reproductive Medicine, Faculty of Medicine, Damascus University, Damascus, Syrian Arab Republic | ||||
Abstract | ||||
Male infertility is a prevalent disorder affecting approximately 7% of the male population, at least 2,000 genes are involved in spermatogenesis, availability of genetic investigations is a challenge in developing countries, GPRC6A, G-protein coupled receptor, activated by Osteocalcin (OCN)an osteoblast secreted hormone, is a master regulator of a complex metabolic and reproductive networks. rs2247911 in GPRC6A plays a role in male fertility and glucose metabolism, Although DNA sequencing is regarded the golden standard tool for genotyping, it is less affordable in developing world laboratories, polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) assay is an inexpensive and highly valuable tool for genotyping, this study aimed to develop (PCR-RFLP) method for assessment of rs2247911 polymorphism.10% of male subjects genotyped previously by sequencing were genotyped the by PCR-RFLP method based on Geneious® bioinformatics tool suggested restriction enzyme (ScrFI), PCR-RFLP 100% correctly identified genotypes. we present for the first time an accurate and inexpensive method for genotyping rs2247911. | ||||
Keywords | ||||
GPRC6A; PCR-RFLP; Sequencing; Osteocalcin; testis failure | ||||
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