Abstract
DNA-based technologies have become widespread tools for soil microbiological analyses in recent years. DNA extraction from the soil is a key step for these approaches: it is a challenge for researchers as it is still both expensive and time-consuming when large surveys are planned. The aim of this study was to develop a high-throughput automated protocol for DNA extraction and purification from soil. The protocol was based on the BioSprint 96 platform and compared for validation with another automated procedure and two commercial column-based kits. To evaluate the performances of the protocols, we considered quality, quantity, and amplifiability of the isolated DNA. The material isolated by means of the four protocols showed appropriate yield and quality and positive amplification. The isolation protocol presented here provided similar results to those of the commercial kits but with two essential differences: cost and time for DNA extraction were drastically reduced. This rapid and efficient protocol is envisaged as ideal to standardize soil studies and treat large numbers of samples, representing a workable alternative to low-throughput and expensive manual extraction methods.
Highlights
Soil is a very complex environment containing huge microbial diversity [1,2]
Information about the chemical-physical properties of the soils analyzed is reported in the Materials and Methods section, together with a description of the isolation methods considered in this work
DNA quantity and purity were measured after DNA isolation with the four protocols as well as their amplifiability
Summary
Soil is a very complex environment containing huge microbial diversity [1,2]. Its characteristics depend on physical and chemical and biological factors [3]. The biotic component forms up to approximately 0.2% of the soil, with microorganisms representing just 20–40% and controlling 80-90%. Of soil processes [4,5]. Biodiversity in soil is enormous, composed of both micro- and mesoorganisms, but the processes in which these are involved are still barely known [6]. Given our poor knowledge concerning the role of the biotic fraction in soil biochemical processes, a deeper understanding of soil biodiversity and its functions is greatly needed. The majority of soil microorganisms cannot be cultivated and characterized by conventional laboratory methods [7,8]. Culture-independent methods are required for their study and DNA-based technologies have been developed over the years to bypass the limits of microorganisms cultivation
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
