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Abstract BackgroundPlant DNA isolation and purification is a time-consuming and laborious process relative to epithelial and viral DNA sample preparation due to the cell wall. The lysis of plant cells to free intracellular DNA normally requires high temperatures, chemical surfactants, and mechanical separation of plant tissue prior to a DNA purification step. Traditional DNA purification methods also do not aid themselves towards fieldwork due to the numerous chemical and bulky equipment requirements. ResultsIn this study, intact plant tissue was coated by hydrophobic magnetic ionic liquids (MILs) and ionic liquids (ILs) and allowed to incubate under static conditions or dispersed in a suspension buffer to facilitate cell disruption and DNA extraction. The DNA-enriched MIL or IL was successfully integrated into the qPCR buffer without inhibiting the reaction. The two aforementioned advantages of ILs and MILs allow plant DNA sample preparation to occur in one minute or less without the aid of elevated temperatures or chemical surfactants that typically inhibit enzymatic amplification methods. MIL or IL-coated plant tissue could be successfully integrated into a qPCR assay without the need for custom enzymes or manual DNA isolation/purification steps that are required for conventional methods. ConclusionsThe limited amount of equipment, chemicals, and time required to disrupt plant cells while simultaneously extracting DNA using MILs makes the described procedure ideal for fieldwork and lab work in low resource environments.
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This content will become publicly available on December 9, 2025
Bacterial Cellulose Purification with Non-Conventional, Biodegradable Surfactants
Bacterial cellulose (BC) is a versatile biopolymer with significant potential across biomedical, food, and industrial applications. To remove bacterial contaminants, such as protein and DNA, BC pellicles undergo purification, which traditionally relies on harsh alkali treatments, such as sodium hydroxide or strong surfactants, which present environmental concerns. In response, this study evaluates the efficacy of various non-conventional surfactants—both non-biodegradable and biodegradable—as alternatives for BC purification. Among the surfactants tested, sodium cocoyl isethionate (SCI), a mild anionic and biodegradable surfactant, emerged as particularly effective, achieving an 80.7% reduction in protein content and a 65.19% reduction in double-stranded DNA (dsDNA) content relative to untreated samples. However, these advantages were not without additional challenges, such as the appearance of residual surfactants. Given SCI’s promising performance and biodegradability, it was further examined in two-step treatment protocols; additionally, sodium dodecyl sulfate (SDS) was also examined as a more traditional anionic surfactant as well as NaOH. For the two-step treatment protocol, BC pellicles were treated with one reagent for 3 h, followed by a second reagent for an additional 3 h. Notably, by using NaOH as the final step in the two-step treatment protocol, residual surfactant was not detected in the FTIR analysis. Overall, this work demonstrates that SCI, in addition to subsequent NaOH treatment, can be used as a surfactant-based approach for BC purification, representing a potential environmentally friendly alternative to traditional surfactant-based approaches for BC purification.
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- Award ID(s):
- 2236940
- PAR ID:
- 10567725
- Publisher / Repository:
- Multidisciplinary Digital Publishing Institute
- Date Published:
- Journal Name:
- Polysaccharides
- Volume:
- 5
- Issue:
- 4
- ISSN:
- 2673-4176
- Page Range / eLocation ID:
- 857 to 871
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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