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- Title
Isolation of nematode DNA from 100 g of soil using Fe<sub>3</sub>O<sub>4</sub> super paramagnetic nanoparticles.
- Authors
GORNY, Adrienne M.; HAY, Frank S.; Xiaohong WANG; PETHYBRIDGE, Sarah J.
- 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, Fe3O4 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.
- Subjects
NEMATODE control; BIOLOGICAL control of nematodes; NORTHERN root-knot nematode; NUCLEIC acid isolation methods; NANOPARTICLES
- Publication
Nematology, 2018, Vol 20, Issue 3, p271
- ISSN
1388-5545
- Publication type
Article
- DOI
10.1163/15685411-00003140