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Prospect and Competence of Quantitative Methods Via Real-Time Pcr in a Comparative Manner: An Experimental Review of Current Methods Publisher



Mahboudi H1 ; Heidari NM2 ; Rashidabadi ZI2 ; Anbarestani AH3 ; Karimi S4 ; Darestani KD5
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Authors Affiliations
  1. 1. Department of medical biotechnology, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
  2. 2. Department of Agronomy and Plant breeding, Agricultural Faculty, Zanjan University, Zanjan, Iran
  3. 3. Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran
  4. 4. Nano Drug Delivery Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran
  5. 5. Biology Department, School of advanced sciences regenerative medicine, Tehran Medical Branch Islamic Azad University, Tehran, Iran

Source: Open Bioinformatics Journal Published:2018


Abstract

Background: There are numerous approaches dealing with relative and absolute quantitation. The methods differ in their efficiency assumption and applicability. Objective: Current methodologies and rations used in qPCR quantification were compared in an experimental study of transgenic copy number determination of a monoclonal antibody Daclizumab. Methods: With an inter and intra-methodical view, variations in relative and absolute quantification strategies were discretely extracted and compared to one another. Results: In relative quantification, six methods were studied and the ratios were computed relative to Glucagon as internal control. For Absolute quantification, the calculations were based on standard curve. Relative quantification considers the relative changes in expression levels while Absolute quantification relates the PCR signal to input copy number with a calibration curve. Conclusion: The observed unevenness of the ratios in Relative approach pointed mainly to the efficiency changes and its calculation formula. Whereas results in Absolute approach strategies showed homogeneity which indicates the consistency of the calculation method. © 2018 Mahboudi et al.
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