A Green and Practical Magnetic Ionic Liquid-Based Microextraction of DNA Using a Low-Cost 3D-Printed Open-Source...

The analysis of nucleic acids offers numerous possibilities such as identification of varied diseases, determination of specific health conditions, and monitoring of the presence of microorganisms. However, DNA extraction remains a critical step that requires cost-effective, time-saving, and efficient experimental workflows that do not hinder PCR amplification. In this study, a novel high-throughput and cost-effective microextraction strategy for DNA isolation using a lab-made open-source 3D-printed magnetic platform was proposed. This affordable multiwell magnetic platform was built using 3D-printed components coupled with electronic modules, capable of promoting z-axis movements using Arduino. In this configuration, 12 magnetic pins were used to hold droplets of the magnetic ionic liquid [P_6,6,6,14 ⁺]-[Ni-(hfacac)₃ ^-], enabling multiple extractions. Only 6.5 μL of the MIL was required for each extraction, and the optimized experimental conditions consisted of 600 μL of sample, extraction time of 25 min, and sample pH at 8, while desorption was performed in 300 μL of Tris-HCl (pH 5.5) for 10 min prior to qPCR analysis. The method exhibited satisfactory performance for capturing the BRAF gene with a linear range from 0.005 to 5 pg/μL, with relative recoveries ranging from 92.2 to 117.0%. Intraday precision ranged from 5.0 to 7.3%, interday precision varied from 5.9 to 11.6%, and the limit of quantification (LOQ) was determined as 0.005 pg/μL. The approach was successfully employed to analyze whole-blood samples, demonstrating its applicability to complex biological matrices. The sustainability and applicability assessments resulted in an SPMS score of 7.58 and a BAGI score of 62.5, indicating that the method is both sustainable and practical. Overall, the low-cost open-source device associated with MIL-based SDME offers an efficient, customizable, and accessible alternative for DNA extraction, with potential for further full automation.