Author:
Mahboudi Hossein,Heidari Negin Mohammadizadeh,Rashidabadi Zahra Irani,Anbarestani Ali Houshmand,Karimi Soroush,Darestani Kaveh Darabi
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.
Publisher
Bentham Science Publishers Ltd.
Subject
Health Informatics,Biomedical Engineering,Computer Science (miscellaneous)
Reference39 articles.
1. Murphy LD, Herzog CE, Rudick JB, Fojo AT, Bates SE.
Use of the polymerase chain reaction in the quantitation of mdr-1 gene expression.
Biochemistry
1990;
29
(45)
: 10351-6.
2. Noonan KE, Beck C, Holzmayer TA, et al.
Quantitative analysis of MDR1 (multidrug resistance) gene expression in human tumors by polymerase chain reaction.
Proc Natl Acad Sci USA
1990;
87
(18)
: 7160-4.
3. Horikoshi T, Danenberg KD, Stadlbauer TH, et al.
Quantitation of thymidylate synthase, dihydrofolate reductase, and DT-diaphorase gene expression in human tumors using the polymerase chain reaction.
Cancer Res
1992;
52
(1)
: 108-16.
4. Heid CA, Stevens J, Livak KJ, Williams PM.
Real time quantitative PCR.
Genome Res
1996;
6
(10)
: 986-94.
5. Winer J, Jung CK, Shackel I, Williams PM.
Development and validation of real-time quantitative reverse transcriptase-polymerase chain reaction for monitoring gene expression in cardiac myocytes in vitro.
Anal Biochem
1999;
270
(1)
: 41-9.
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