A lysis-free, all-in-one platform based on oligonucleotide-functionalized magnetic ionic liquid for direct and...

The extraction of DNA from plant tissues is often hindered by the resilient cell wall, which necessitates cumbersome pretreatment steps in conventional methods. Furthermore, existing techniques typically co-extract total DNA without the capability for sequence-specific isolation. To address these limitations, this study developed a novel oligonucleotide-functionalized hydrophobic magnetic ionic liquid (oligo-HMIL). This material integrates high stability, strong magnetic responsiveness, and exceptional hydrophobicity, allowing it to be magnetically manipulated to directly penetrate plant cell walls and membranes for DNA extraction, thereby eliminating the need for a separate lysis step. More significantly, the material enables sequence-specific recognition and selectively isolates the target DNA from complex matrices containing substantial amounts of non-target DNA, proteins, and other interferents. Based on this material, a lysis-free, all-in-one platform for direct and sequence-specific extraction of DNA was established. Under optimized conditions, the method achieved efficient extraction and enrichment of the target DNA from various plant tissues, with an extraction efficiency of 78.25 ± 1.83%. Compared to a commercial kit, the proposed approach increased DNA yield by at least 3-fold while reducing the processing time from 4 h to 30 min and simplifying the workflow. The material can be recycled and reused, and the method demonstrated excellent stability (RSD = %, n = 3) and reproducibility (RSD = 2.34%, n = 3). This work provides a green, rapid, and reusable platform for targeted plant DNA extraction, holding broad application potential in genetic research and biotechnology.