Study of the Effect of Material of Microfluid Chip on the Polymerase Chain Reaction
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DIAGNOSTICS AND PROBE TECHNOLOGIES IN BIOLOGY AND MEDICINE
Study of the Effect of Material of Microfluid Chip on the Polymerase Chain Reaction A. N. Tupika,*, G. E. Rudnitskayaa, T. A. Lukashenkoa, and A. A. Evstrapova aInstitute
for Analytical Instrumentation, Russian Academy of Sciences, St. Petersburg, 190103 Russia *e-mail: [email protected] Received December 26, 2019; revised December 26, 2019; accepted February 17, 2020
Abstract—Effect of several polymer materials from different manufacturers on the polymerase chain reaction is tested. Absence of adsorption of reaction components at room temperature is demonstrated for a polymethylmethacrylate chip. However, a decrease in the reaction efficiency at low DNA concentrations is observed for the polymerase chain reaction in the microchip. DOI: 10.1134/S1063784220090261
INTRODUCTION Microfluid chips (microchips) contain a system of micro-sized channels and reservoirs. A relatively small volume makes it possible to decrease amounts of reagents, arrange a large number of reaction chambers on a single device, and implement new analytical approaches and procedures. Microchips are widely used in genetic analysis to develop compact automated systems for complex analysis of samples [1–4]. Polymerase chain reaction (PCR) is used to obtain amounts of DNA fragments that are sufficient for detection, identification, and analysis of nucleic acids. A specific feature of the PCR is related to multiple cyclic heating and cooling. Miniaturization provides efficient conditions for reactions and a decrease in the reaction time. However, relatively high ratio of the surface area to the volume of microstructures (from 3–10 to ~150 in comparison with 1.4 for tubes [5]) leads to a stronger effect of the working surface on the results of analysis due to contact of material and biological sample and reagents. Such an effect is related to both chemical interaction and physical adsorption of mixture components on the inner surface of microstructures. Adsorption may result in variations in the ratio of components in solution and, hence, a decrease in the efficiency of reaction up to it inhibition. Thus, protective coatings (static modification) [6, 7] or surfactants that compete with reaction components in adsorption on the microchip walls (dynamic modification) [8, 9] are used in the PCR chips. A combination of the two methods (e.g., preliminary washing of microstructures using a solution of bovine serum albumin (BSA)) [5, 10] is also possible. An increase in the concentration of adsorbed reagents leads to sufficient amounts of reagents in the reaction mixture (a concentration of 0.1 units/μL can be used instead of
0.025 units/μL for silicon microchips [8]) but the analysis becomes impossible at small concentrations of DNA upon adsorption of nucleic acids. Most widely used materials for microchips (silicon, quartz glasses, polycarbonate (PC), polymethylmethacrylate (PMMA), etc. [11]) are well studied and the corresponding technologies for fabrication of microstructures are developed. Mater
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