en بیوانفورماتیک Bioinformatic پژوهشي Research <span style="font-size:11pt"><span style="line-height:107%"><span calibri="" style="font-family:"><b><span style="font-size:12.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">Background: </span></span></span></b><span style="font-size:12.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">Designing the primer pairs is one of the most important factors in 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 with minimum expression. To achieve this aim, we selected two candidate genes with little expression, namely, G-protein coupled receptor 120 (<i>GPR120</i>) and peroxisome proliferator-activated receptor-&gamma; (<i>PPAR&gamma;</i>), in peripheral blood leukocytes of healthy volunteers.</span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:107%"><span calibri="" style="font-family:"><b><span style="font-size:12.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">Methods: </span></span></span></b><span style="font-size:12.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">Peripheral blood was collected from 30 healthy volunteers. Primers for <i>GPR120 </i>and <i>PPAR&gamma; </i>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 manufacturer instructions. Finally, the melting curve analysis of <i>GPR120 </i>and <i>PPAR&gamma; </i>was assessed using the quantitative real-time PCR method.</span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:107%"><span calibri="" style="font-family:"><b><span style="font-size:12.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">Results: </span></span></span></b><span style="font-size:12.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">The in silico gene expression investigation revealed that <i>GPR120 </i>and <i>PPAR&gamma; </i>have minimal leukocyte expression. Besides, the melting curves analysis for both genes in the studied individuals showed only one melting peak, confirming the specific amplification of the desired genes.</span></span></span></span></span></span><br> <span style="font-size:11pt"><span style="line-height:107%"><span calibri="" style="font-family:"><b><span style="font-size:12.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">Conclusion: </span></span></span></b><span style="font-size:12.0pt"><span style="line-height:107%"><span new="" roman="" style="font-family:" times="">Altogether, the study findings indicated that we could utilize the peripheral blood sample for assessing the gene expression and amplification of omega-3 fatty acids receptors, i.e. <i>GPR120 </i>and <i>PPAR&gamma; </i>as two candidate genes with very low expression in leukocytes.</span></span></span></span></span></span><br> &nbsp; GPR120, Omega-3 fatty acid, cDNA, PPARγ, Real-time PCR 205 214 http://rmm.mazums.ac.ir/browse.php?a_code=A-10-1222-3&slc_lang=en&sid=1 Ramin Lotfi ramin.lotfi1370@gmail.com 100319475328460011614 100319475328460011614 Yes Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Kurdistan Regional Blood Transfusion Center, Sanandaj, Iran Farhad Salari f.salari@kums.ac.ir 100319475328460011615 100319475328460011615 No Department of Immunology, School of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran