Abstract
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 (A260/A280), making it suitable for various applications aiming at nucleic acid detection.
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All data supporting the findings of this study are available within the paper and its Supplementary Information. All images in this study were created by the authors and are original designs. There has been no use of images sourced from external locations, ensuring compliance with copyright restrictions.
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Acknowledgements
The authors thank the AICTE - IDEA (Idea Development, Evaluation & Application) Lab and the Central Instrumentation Facility, BIT Mesra, for the technical and instrumentation support.
Funding
D.P. has received grant from the parent Institute for this work. (Ref: BT/HOD/22–23/074). R.G. received fellowship (APO/PhD/IRS/2023–2024/61).
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Conceptualization: DP; Methodology: RG, AM, NKM, DP; Formal analysis and investigation: RG, DP, Writing—original draft preparation: RG, AM; Writing—review and editing: DP, NKM; Funding acquisition: DP; Resources: DP; Supervision: DP, NKM.
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Garg, R., Maurya, A., Mani, N.K. et al. Thread-powered cell lysis and isotachophoresis: unlocking microbial DNA for diverse molecular applications. World J Microbiol Biotechnol 40, 97 (2024). https://doi.org/10.1007/s11274-024-03906-2
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DOI: https://doi.org/10.1007/s11274-024-03906-2