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UNIVERSITY OF BUCHAREST
FACULTY OF PHYSICS Guest
2024-11-23 17:50 |
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Conference: Bucharest University Faculty of Physics 2014 Meeting
Section: Biophysics; Medical Physics
Title:
A comparative analysis of three methods used for RNA concentration assay
Authors:
Sorina SCHIPOR(1), Suzana VLADOIU(1), Ancuta Elena BACIU(2,1), Ana Maria NICULESCU(3), Andra CARAGHEORGHEOPOL(1),
Iulia IANCU(4), Adriana PLESA(4), A. I. POPESCU(2), Dana MANDA (1)
Affiliation:
(1) “C. I. Parhon” National Institute of Endocrinology, Bucharest, Romania
(2) University of Bucharest, Faculty of Physics, P.O. Box MG-11, Magurele, Romania
(3) Agilrom Scientific, Bucharest, Romania
(4) “Stefan S. Nicolau” Institute of Virology, Bucharest, Romania
E-mail
ancutaelenabaciu@yahoo.com
Keywords:
RNA quantitation, UV spectrophotometry, fluorimetry, microfluidic capillary electrophoresis.
Abstract:
High RNA sample quality is essential for a successful gene expression and microarray experiments. Two simultaneous conditions should be accomplished by the RNA samples: the structural integrity of the molecules and an adequate concentration.The objective of this study is to do a comparative analysis between three different methods of measuring RNA concentration.Methods. The three methods that are considered here are: UV spectrophotometry, spectrophotofluorimetry, and microfluidic capillary electrophoresis. Sixteen RNA samples were assayed by these three methods.Results. The principles and the results of each method as well as advantages, disadvantages and perturbations factors are analyzed and discussed. It was observed that the measured concentrations are correlated to each other. Fluorescent labelling of the RNA gives more accurate results even in the presence of the frequent contaminants like DNA and proteins.Conclusions. Knowing the strength and the limits of the methods required for RNA quantitation, the scientist can choose the most cost-effective protocol.
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