Comparison of SYBR Green and TaqMan methods in quantitative real-time polymerase chain reaction analysis of four adenosine receptor subtypes

Authors
Mohamadhasan Tajadini 1
Mojtaba Panjehpour 2
Shaghayegh Haghjooy Javanmard 3
1 Department of Clinical Biochemistry, School of Pharmacy and Pharmaceutical Research Center; Physiology Research Center, Isfahan University of Medical Sciences, Isfahan, Iran
2 Department of Clinical Biochemistry, School of Pharmacy and Pharmaceutical Research Center, Isfahan University of Medical Sciences, Isfahan, Iran
3 Physiology Research Center, Isfahan University of Medical Sciences, Isfahan, Iran
Abstract
Background: Real-time polymerase chain reaction (PCR) is based on the revolutionary method of PCR. This technique is the result of PCR enormous sensitivity and real-time monitoring combination. In quantitative gene expression analysis, two methods have more popularity, SYBR Green and TaqMan, SYBR Green is relatively cost benefit and easy to use and technically based on binding the fluorescent dye to double-stranded deoxyribonucleic acid (dsDNA) where TaqMan method has more expensive and based on dual labeled oligonucleotide and exonuclease activity of Taq polymerase enzyme. Specificity is the most important concern with the usage of any non-specific dsDNA-binding Dyes such as SYBR Green whiles more specificity showed by labeled oligonucleotide method such as TaqMan. In this study, we compared two common RT PCR methods, TaqMan and SYBR Green in measurement gene expression profile of adenosine receptors.
Materials and Methods: Gene expression profiles of A1, A2A, A2B and A3 Adenosine receptors were analyzed by optimized TaqMan and SYBR Green quantitative RT PCR in breast cancer tissues. Primary expression data was normalizing by B. actin reference gene.
Results: Efficiencies were calculated more than 95% for TaqMan and SYBR Green methods in all genes. The correlations between means of normalized data of each gene in two methods were positive and significant (P < 0.05).
Conclusion: Data analysis showed that with the use of high performance primer and by use proper protocols and material we can make precise data by SYBR Green as TaqMan method. In other word by optimization of SYBR Green method, its performance and quality could be comparable to TaqMan method.
Keywords
Quantitative real-time polymerase chain reaction
SYBR Green
TaqMan
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Advanced Biomedical Research
Volume 2014, February
February 2014
Pages 1-6