Thread-powered cell lysis and isotachophoresis: unlocking microbial DNA for diverse molecular applications

• Published in: World journal of microbiology & biotechnology Vol. 40; no. 3; p. 97
• Main Authors: Garg, Rishabh, Maurya, Aharnish, Mani, Naresh Kumar, Prasad, Dinesh
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer Netherlands 01.03.2024; Springer Nature B.V
• Subjects: Applied Microbiology; Biochemistry; Biomedical and Life Sciences; Biotechnology; Blue Light; Cell Death; Cell walls; Deoxyribonucleic acid; DNA; Electric fields; Electrolytes; Environmental Engineering/Biotechnology; Ion migration; Isotachophoresis; Life Sciences; Lysis; Microbiology; Microorganisms; Molecular biology; Nucleic acids; Polypropylene; Polypropylenes; Purification; DNA extraction; Microbial DNA purification; DNA isoalation; Thread isotachophoresis; PCR inhibitors
• ISSN: 0959-3993; 1573-0972
• DOI: 10.1007/s11274-024-03906-2

Central to the domain of molecular biology resides the foundational process of DNA extraction and purification, a cornerstone underpinning a myriad of pivotal applications. In this research, we introduce a DNA extraction and purification technique leveraging polypropylene (PP) threads. The process commences with robust cell lysis achieved through the vigorous agitation of interwoven PP threads. The friction between the threads facilitates cell lysis especially those microbes having tough cell wall. For purification of DNA, thread-based isotachophoresis was employed which makes the whole process swift and cost-effective. Lysed cell-laden threads were submerged in a trailing electrolyte which separated DNA from other cellular contents. The process was performed with a tailored ITP device. An electric field directs DNA, cell debris, trailing electrolyte, and leading electrolyte toward the anode. Distinct ion migration resulted in DNA concentrating on the PP thread’s anode-proximal region. The SYBR green dye is used to visualize DNA as a prominent green zone under blue light. The purified DNA exhibits high purity levels of 1.82 ± 0.1 (A 260 /A 280 ), making it suitable for various applications aiming at nucleic acid detection.