Fast and Sustainable Thermo-osmotic DNA Extraction Protocol for Trans-spectrum Contingency and Field Use.

Area of Science:

• Molecular Genetics
• Biotechnology

Background:

• Standard DNA extraction kits are often proprietary and sample-specific, hindering resource sharing during supply shortages.
• Existing rapid extraction methods have limitations, necessitating simpler, more adaptable protocols.

Purpose of the Study:

• To develop a cost-effective and accessible DNA extraction protocol using boiling water for rapid molecular diagnostics.
• To address limitations of current DNA extraction methods, particularly during crises or in resource-limited settings.

Main Methods:

• Utilizing boiling-hot water to lyse cells, virions, and nuclei from various biosamples.
• Employing serial dilutions of the crude extract to optimize DNA and inhibitor concentrations for direct PCR.
• Performing direct PCR on crude lysate and dilutions without purification or quantification steps.

Main Results:

• Successfully extracted DNA from uncomplicated, enriched samples like fruit mycelia and human swabs using boiling water.
• Demonstrated the feasibility of direct PCR with crude extracts and dilutions, bypassing purification.
• Achieved cost reduction and a lower environmental footprint compared to traditional DNA extraction methods.

Conclusions:

• Boiling water DNA extraction is a viable, low-cost alternative for rapid molecular testing, especially in resource-limited or crisis situations.
• This method enhances the sustainability of field operations and standard laboratory practices facing supply chain disruptions.