Nowadays, there is an increasing demand for fast sustainable, and reliable plant biomolecular analyses. Plant DNA can be used for several fast screening applications including, for instance, DNA barcoding for food traceability [1]. In general, DNA isolation requires multiple and time-consuming steps to remove cellular interferences with the consequence that sample preparation is the main bottleneck in a bioanalytical workflow. New perspectives have been opened by the use of Magnetic Ionic Liquids (MILs), proposed in 2015 by Anderson’s group, as new materials for selective extraction and purification of nucleic acids [2]. Hydrophobic MILs can be used in magnet-assisted Dispersive Liquid-Liquid Microextraction (maDLLME) by dispersing the MILs into very small microdroplets in a water solution containing the nucleic acids and by recovering the MILs with the application of a magnetic field. This approach results in reduced extraction time and increased extraction efficiency toward nucleic acids. So far, it has been applied to DNA (purified or in complex solutions containing proteins) and to bacterial cell lysate, but, to date, no applications on plants have been reported [3]. In the present study, three low-viscosity hydrophobic MILs, containing Co, Mn and Ni [4], were tested for the isolation of plant DNA. Preliminary tests were carried out on purified DNA from Arabidopsis thaliana (L.) Heynh. while subsequent experiments were performed directly on the plant material. The isolation of the DNA was performed by maDLLME after lysis of the plant cells and the DNA was then recovered by back extraction. The isolated DNA was subjected to qualitative and quantitative analyses, with interesting results in terms of MILs-based method extraction performance. In the field of plant molecular biology, the application of maDLLME with MILs could represent a powerful tool for fast, sustainable and potentially automatable screening methods involving DNA.
Exploiting Magnetic Ionic Liquids to Develop a One-step Method for Plant DNA Isolation
Cecilia Cagliero;Arianna Marengo;Carlo Bicchi;Cinzia Bertea;Patrizia Rubiolo
2018-01-01
Abstract
Nowadays, there is an increasing demand for fast sustainable, and reliable plant biomolecular analyses. Plant DNA can be used for several fast screening applications including, for instance, DNA barcoding for food traceability [1]. In general, DNA isolation requires multiple and time-consuming steps to remove cellular interferences with the consequence that sample preparation is the main bottleneck in a bioanalytical workflow. New perspectives have been opened by the use of Magnetic Ionic Liquids (MILs), proposed in 2015 by Anderson’s group, as new materials for selective extraction and purification of nucleic acids [2]. Hydrophobic MILs can be used in magnet-assisted Dispersive Liquid-Liquid Microextraction (maDLLME) by dispersing the MILs into very small microdroplets in a water solution containing the nucleic acids and by recovering the MILs with the application of a magnetic field. This approach results in reduced extraction time and increased extraction efficiency toward nucleic acids. So far, it has been applied to DNA (purified or in complex solutions containing proteins) and to bacterial cell lysate, but, to date, no applications on plants have been reported [3]. In the present study, three low-viscosity hydrophobic MILs, containing Co, Mn and Ni [4], were tested for the isolation of plant DNA. Preliminary tests were carried out on purified DNA from Arabidopsis thaliana (L.) Heynh. while subsequent experiments were performed directly on the plant material. The isolation of the DNA was performed by maDLLME after lysis of the plant cells and the DNA was then recovered by back extraction. The isolated DNA was subjected to qualitative and quantitative analyses, with interesting results in terms of MILs-based method extraction performance. In the field of plant molecular biology, the application of maDLLME with MILs could represent a powerful tool for fast, sustainable and potentially automatable screening methods involving DNA.
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