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TITLE:

MOLECULAR BASED DETECTION OF A PIK3C3 (PHOSPHATIDYLINOSITOL 3-KINASE) GENE VARIATIONS IN T2D PATIENTS OF TABUK-SAUDI ARABIA .

AUTHORS:

Mohammed Sakran, Fahad M Almutairi, Adel I.A. AL-Alawy, Rashid Mir, and Imadeldin Elfaki*

ABSTRACT:

Background: The PI3K pathway is commonly dysregulated in different diseases such as cancer, with the most common events being mutation or increased gene copy numbers of PIK3CA or other PI3K isoforms, loss of expression of the pathway suppressors (for example, PTEN), or hyperactivation of RTKs through receptor overexpression or activating mutations. Several gene polymorphisms have been frequently associated with the onset of T2D. Therefore the aim of this study was to optimize molecular based assay for the detection of PIK3C rs121913281 C>T gene variation in T2D patients. Methodology: This study was conducted on 60 samples among which 30 were T2D patients and 30 were healthy controls. DNA was extracted by Qiagen Kit and AS-PCR system was optimized to detect PIK3C rs121913281 C>T gene variation in T2D patients. Results: This study was conducted on 60 samples among which 30 were T2D patients and 30 were healthy controls. The 30 newly diagnosed T2D patients. AS-PCR system was optimized to detect PIK3C rs121913281 C>T gene variation in clinically confirmed cases of T2D patients and healthy controls. The technique was successfully optimized by using wild-type or mutant-type primers with matched or one-base mismatched to examine the known SNPS in PIK3C rs121913281 C>T. Result indicated that AS-PCR technique can be used as a potential molecular tool in the detection of potential PIK3C gene variations in T2Ds.Conclusion: Allele Specific-PCR for PIK3CA- rs121913281 C>T gene variation was successfully optimized. The assay proved to be fast, accurate, simple and economical that does not entail any special equipment other than a thermocycler and gel documentation system. Key Words: Type 2 diabetes (T2D), Allele-specific PCR, PIK3CA, Phosphatidylinositol 3-kinases (PI3Ks), rs121913281 C>T

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