Skip to main content
Article
Development of an innovative and sustainable one-step method for rapid plant DNA isolation for targeted PCR using magnetic ionic liquids
Plant Methods
  • Arianna Marengo, Università di Torino
  • Cecilia Cagliero, Università di Torino
  • Barbara Sgorbini, Università di Torino
  • Jared L. Anderson, Iowa State University
  • Miranda N. Emaus, Iowa State University
  • Carlo Bicchi, Università di Torino
  • Cinzia M. Bertea, Università di Torino
  • Patrizia Rubiolo, Università di Torino
Document Type
Article
Publication Version
Published Version
Publication Date
3-9-2019
DOI
10.1186/s13007-019-0408-x
Abstract

Background

Nowadays, there is an increasing demand for fast and reliable plant biomolecular analyses. Conventional methods for the isolation of nucleic acids are time-consuming and require multiple and often non-automatable steps to remove cellular interferences, with consequence that sample preparation is the major bottleneck in the bioanalytical workflow. New opportunities have been created by the use of magnetic ionic liquids (MILs) thanks to their affinity for nucleic acids. Results

In the present study, a MIL-based magnet-assisted dispersive liquid–liquid microextraction (maDLLME) method was optimized for the extraction of genomic DNA from Arabidopsis thaliana (L.) Heynh leaves. MILs containing different metal centers were tested and the extraction method was optimized in terms of MIL volume and extraction time for purified DNA and crude lysates. The proposed approach yielded good extraction efficiency and is compatible with both quantitative analysis through fluorimetric-based detection and qualitative analysis as PCR amplification of multi and single locus genes. The protocol was successfully applied to a set of plant species and tissues. Conclusions

The developed MIL-based maDLLME approach exhibits good enrichment of nucleic acids for extraction of template suitable for targeted PCR; it is very fast, sustainable and potentially automatable thereby representing a powerful tool for screening plants rapidly using DNA-based methods.

Comments

This article is published as Marengo, Arianna, Cecilia Cagliero, Barbara Sgorbini, Jared L. Anderson, Miranda N. Emaus, Carlo Bicchi, Cinzia M. Bertea, and Patrizia Rubiolo. "Development of an innovative and sustainable one-step method for rapid plant DNA isolation for targeted PCR using magnetic ionic liquids." Plant Methods 15 (2019): 23. DOI: 10.1186/s13007-019-0408-x. Posted with permission.

Creative Commons License
Creative Commons Attribution 4.0
Copyright Owner
The Authors
Language
en
File Format
application/pdf
Citation Information
Arianna Marengo, Cecilia Cagliero, Barbara Sgorbini, Jared L. Anderson, et al.. "Development of an innovative and sustainable one-step method for rapid plant DNA isolation for targeted PCR using magnetic ionic liquids" Plant Methods Vol. 15 (2019) p. 23
Available at: http://works.bepress.com/jared_anderson/28/