Isolation of nematode DNA from 100 g of soil using Fe3O4 super paramagnetic nanoparticles

Abstract

An economical method for extracting nematode DNA from 100 g of soil was developed to facilitate nematode detection and quantification, and tested using the Northern root-knot nematode, Meloidogyne hapla . The method utilised enzymatic laundry detergent lysis, Fe 3 O 4 super paramagnetic iron oxide nanoparticle (SPION) capture, and polyvinylpolypyrrolidone (PVPP) purification. Resultant DNA from this SPION capture method was approximately 100-fold less but of similar quality to DNA obtained from a standard phenol procedure and a commercial DNA extraction kit. An addition of 10 mg of nanoparticles to the extraction lysate was identified to maximise DNA yield while minimising co-capture of contaminants. The detection limit of the SPION capture method was approximately 100 nematodes (100 g soil) −1 . The SPION capture method extracted nematode DNA from mineral soils but requires further optimisation for extraction from high organic matter ( i.e. , ‘muck’) soils. The benefits of this method compared to alternative techniques are discussed.