Solid-Phase PCR on Film Biochips with Brush Polymer Cells, "Lab-on-a-Chip"

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Abstract

A method of nucleic acid analysis by solid-phase PCR with immobilised primers extension (primer extension) in a closed film biochip during thermocycling, with control of Cy5-fluorescence-labelled nucleotide incorporation by digital fluorescence microscopy, by endpoint, was developed. A "film biochip", made of film polyethylene terephthalate, with an internal chamber, with brush polymer cells with immobilised primers, with channels for supply and removal of solutions into the biochip chamber, has been developed, with low heat capacity and high thermal conductivity of thin film components of the biochip, with thermocycling, with registration of results by digital fluorescence microscopy through the lid and the layer of washing liquid without disassembling the biochip, in a system isolated from the environment, "laboratory on a chip". The performance of the method and functional suitability of the "film biochip" was demonstrated by analysing samples containing DNA from pathogenic bacteria Staphylococcus auereus and Legionella pneumophila.

About the authors

I. Yu Shishkin

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Russia, Moscow

K. A Simikov

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Russia, Moscow

G. F Shrylev

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Russia, Moscow

R. A Miftakhov

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Russia, Moscow

O. A Zasedateleva

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Russia, Moscow

V. E Kuznetsova

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Russia, Moscow

V. E Shershov

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Russia, Moscow

S. A Surzhikov

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Russia, Moscow

V. A Vasiliskov

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Russia, Moscow

S. A Lapa

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Russia, Moscow

A. V Chudinov

Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: chudhome@rambler.ru
Russia, Moscow

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