SPION-mediated soil DNA extraction and comparative analysis with conventional and commercial kit-based protocol.
Direct isolation of soil DNA comes as an emerging technology to understand the microbial diversity of a particular environment circumventing the dependency on culturable methods. Soil DNA isolation is tough due to the presence of various organic components present in soil which interfere in extraction procedure. Here, we report a novel direct soil DNA extraction protocol utilizing bare superparamagnetic iron oxide nanoparticles and its comparison with conventional and commercial kit-based soil DNA extraction methods. The quality, quantity and feasibility of the recovered DNA from all the three methods towards various molecular techniques were checked. Our magnetic nanoparticle-based soil DNA extraction successfully yields pure DNA without any RNA or protein contamination as revealed by the nanodrop spectrophotometer and agarose gel electrophoretic study. Different methods of soil DNA extraction were evaluated on the basis of PCR, denaturing gradient gel electrophoresis and real-time PCR. Soil DNA extracted using conventional method fails to carry out critical molecular biology techniques where as magnetic nanoparticle-based soil DNA extraction gave good results which is comparable to commercial kit. This comparative study suggests that protocol described in this report is novel, less time consuming, cost effective with fewer handling steps and yields high quantity, good quality DNA from soil.
Department of Microbiology, University of Kalyani, Nadia, West Bengal, India.
Tanima
T
Paul
Department of Microbiology, University of Kalyani, Nadia, West Bengal, India.
Semanti
S
Basu
Department of Microbiology, University of Kalyani, Nadia, West Bengal, India. drkekasarkar@yahoo.com.
Keka
K
Sarkar
16S rDNA; Density gradient gel electrophoresis (DGGE); Real-time PCR; Soil DNA extraction; Superparamagnetic iron oxide nanoparticle (SPION)