International Conference PhysicA.SPb/2023

Optimization of the lysis process for flow-through isolation of nucleic acids

M. V. Zaitseva, I. E. Antifeev, D. G. Petrov, N. A. Esikova, E. D. Makarova

Institute for Analytical Instrumentation, Russian Academy of Sciences, Saint Petersburg, Russia

Abstract: The influence of the chemical lysis time and temperature on the results of E. coli DNA extraction was studied in order to reduce the time without losing quality of nucleic acid isolation. Values of threshold cycles (Ct) of the studied samples obtained by extracting DNA from E. coli bacteria were determined and analyzed at various lysis parameters: time (5, 7, 9, 11, 13 and 15min) and temperature (+45, +55, +65, and +75$^\circ$). It has been shown that it is possible to reduce the lysis time by 67% (10 min) and cell lysis process temperature by 15% (10$^\circ$C) without losing the efficiency as compared with the recommended conditions (15 min, 65$^\circ$C).

Keywords: PCR, sample preparation, lysis, temperature, time.

Received: 19.05.2023
Revised: 26.06.2023
Accepted: 30.10.2023

DOI: 10.61011/PJTF.2023.23.56847.176A

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