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Research in Molecular Medicine، جلد ۱۰، شماره ۴، صفحات ۰-۰
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عنوان انگلیسی Designing Specific Primers for Amplification and Quantitative Analysis of the GPR120 and PPARγ Genes
چکیده انگلیسی مقاله Background: Designing the primer pairs is one of the most important factors affecting the amplification and quantitative analysis of the nucleic acid sequences of interest. Using in silico methods, the present study intends to design highly specific primers for quantitative analysis of the genes having very low expression. To achieve this aim, we selected two candidate genes having very low expression, namely, G-protein coupled receptor 120 (GPR120) and peroxisome proliferator-activated receptor-g (PPARg), in peripheral blood leukocytes of healthy volunteers.
Materials And Methods Peripheral blood was collected from 30 healthy volunteers. Primers for GPR120 and PPARγ were designed using online websites (UCSC, Oligocalc, and Oligoanalyzer) and the primer designing tool (NCBI). Total RNA extraction and cDNA synthesis were done using commercially available kits based on the instructions of the manufacturers. Finally, the melting curve analysis of GPR120 and PPARγ was assessed using the quantitative real-time PCR method.
Results: The investigation of the in silico gene expression revealed that GPR120 and PPARγ have very low expression in leukocytes. Besides, the melting curves analysis for GPR120 and PPARg genes in the studied individuals showed only one melting peak, confirming the specific amplification of the desired genes.
Conclusions: Altogether, the study findings indicated that we can utilize the peripheral blood sample for assessing the gene expression and amplification of omega-3 fatty acids receptors, i.e. GPR120 and PPARg as two candidate genes possessing very low expression in leukocytes.
کلیدواژه‌های انگلیسی مقاله GPR120, Omega-3 fatty acid, cDNA, PPARγ, Real-time PCR
نویسندگان مقاله | Ramin Lotfi
Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Kurdistan Regional Blood Transfusion Center, Sanandaj, Iran


| Farhad Salari
Department of Immunology, School of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran
نشانی اینترنتی http://rmm.mazums.ac.ir/browse.php?a_code=A-10-1222-3&slc_lang=en&sid=1
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زبان مقاله منتشر شده en
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نوع مقاله منتشر شده پژوهشی
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