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Mycobacterial DNA Extraction using Bead Beating in Custom Buffer Followed by NGS Workflow
Published on: June 13, 2025
High-throughput Method of One-Step DNA Isolation for PCR Diagnostics of Mycobacterium tuberculosis
D V Kapustin1, A I Prostyakova1, Ya I Alexeev2
1M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, GSP-7, Ul. Miklukho-Maklaya, 16/10, Moscow, Russia, 117997.
Abstract:
The efficiency of one-step and multi-step protocols of DNA isolation from lysed sputum samples containing the Mycobacterium tuberculosis complex has been compared. DNA was isolated using spin-cartridges containing a special silica-based sorbent modified with fluoroplast and polyaniline, or using an automated isolation system. One-step isolation using the obtained sorbent has been shown to ensure a significantly lower DNA loss and higher sensitivity in the PCR detection of Mycobacterium tuberculosis as compared to a system based on sorption and desorption of nucleic acids during the isolation.

