Evaluation of Real-Time PCR Efficiency by the Use of Two Strategies: Standard Curve and Linear Regression

Document Type : Original Article

Authors

1 Assistant Professor of Immunology.Department of Immunology, & Cellular & Molecular Research Center, School of Medicine, Ahvaz, Jundishapur University of Medical Sciences, Ahvaz, Iran.

2 Master of Science Student.Department of Immunology, & Cellular & Molecular Research Center, School of Medicine, Ahvaz, Jundishapur University of Medical Sciences, Ahvaz, Iran.

3 Associated Professor of Immunology. Department of Immunology, School of Medicine, Ahvaz, Jundishapur University of Medical Sciences, Ahvaz, Iran.

4 Assistant Professor of Medical Genetics. Department of Medical Genetics & Cellular & Molecular Research Center, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.

Abstract

Background and Objective: The detection of nucleic acids using Real-Time CPR has many application .Quantitative analysis of mRNA using Real-Time PCR by Relative and Absolute methods. Widely used in biological studies. The purpose of this study was to compare the Relative and calculated PCR Efficiency by standard curve and LinRegPCR methods.
Subject and Methods: After sampling and extraction of RNA and cDNA synthesis,the quantitative RT-PCR was performed, then the PCR Efficiency was calculated by Using the two methods of standard curve and LinRegPCR. At the end the results of the two methods were compared and analyzed.
Results: The efficiency of PCR for the GAPDH, TGF-β and IL-10 genes with the standard curve method were 1.99, 1.81 and 1.87, respectively. The PCR efficiency of these three genes were 1.98, 1.82 and 1.82 by using LinRegPCR software method, respectively.
Conclusion: The analysis of the data obtained from PCR proliferation of cDNA of the three genes, GAPDH, TGF β and IL-10 showed no statistical difference between standard curve and LineRegPCR methods. Therefore, it is recommended to use the LinRegPCR software method instead of the expensive standard curve method.                              

Keywords

References

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Jundishapur Scientific Medical Journal
Volume 11, Issue 1 - Serial Number 76
July and August 2002
Pages 85-95

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History

  • Receive Date: 04 October 2011
  • Revise Date: 26 November 2011
  • Accept Date: 21 December 2011

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