PCR-ready human DNA extraction from urine samples using magnetic nanoparticles

Urine-derived human genomic DNA (gDNA) has wide application in a variety of disciplines including clinical medicine, sports, and forensic science. We describe a novel method for gDNA extraction from urine samples using carboxylated magnetic nanoparticles (CMNPs) as solid-phase adsorbents. Sedimentation associated with freezing of urine samples significantly reduces cell capture by CMNPs. However, the addition of 10 mM EDTA and subsequent pH modification (pH 6.0–7.1) can re-dissolve urine sediments. Purified gDNA ranged from around 0.1 kb to more than 23 kb. PCR using specific primers targeting K-ras, GAPDH, CYP3A4 and GDF5 amplified 100% of varying sized gene fragments, verifying the high quality of the isolated DNA. Successful PCR amplifications using DNA isolated from urine samples as small as 50 μl were demonstrated. Enrichment of urine cells and subsequent adsorption of DNA can be achieved with the same CMNPs, greatly simplifying extraction procedures. The CMNP gDNA extraction technique proved to be simple, rapid, sensitive and environmentally friendly, with application for routine laboratory use and potentially within automated urine extraction platforms.

► A method for urine DNA extraction using carboxylated magnetic nanoparticles was developed.
► The addition of 10 mM EDTA and pH modification (pH 6.0–7.1) can re-dissolve urine sediments.
► Purified DNA ranged from around 0.1 kb to more than 23 kb.
► DNA quality was validated by its yield, molecular weight, and the ability to serve as PCR templates.
► The developed method proved to be simple, rapid, sensitive and environmentally friendly.