Direct extraction of genomic DNA from maize with aqueous ionic liquid buffer systems for applications in genetically modified organisms analysis.
To date, the extraction of genomic DNA is considered a bottleneck in the process of genetically modified organisms (GMOs) detection. Conventional DNA isolation methods are associated with long extraction times and multiple pipetting and centrifugation steps, which makes the entire procedure not only tedious and complicated but also prone to sample cross-contamination. In recent times, ionic liquids have emerged as innovative solvents for biomass processing, due to their outstanding properties for dissolution of biomass and biopolymers. In this study, a novel, easily applicable, and time-efficient method for the direct extraction of genomic DNA from biomass based on aqueous-ionic liquid solutions was developed. The straightforward protocol relies on extraction of maize in a 10 % solution of ionic liquids in aqueous phosphate buffer for 5 min at room temperature, followed by a denaturation step at 95 °C for 10 min and a simple filtration to remove residual biopolymers. A set of 22 ionic liquids was tested in a buffer system and 1-ethyl-3-methylimidazolium dimethylphosphate, as well as the environmentally benign choline formate, were identified as ideal candidates. With this strategy, the quality of the genomic DNA extracted was significantly improved and the extraction protocol was notably simplified compared with a well-established method.
IFA-Tulln, Center of Analytical Chemistry, Vienna University of Technology, Konrad Lorenz Str. 20, 3430, Tulln, Austria.
Eric
E
Gonzalez García
Anna K
AK
Ressmann
Peter
P
Gaertner
Ronald
R
Zirbs
Robert L
RL
Mach
Rudolf
R
Krska
Katharina
K
Bica
Kurt
K
Brunner
D002021:Buffers; D000077286:Cetrimonium; D002593:Cetrimonium Compounds / Q000737:chemistry; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification*; D018745:Genome, Plant; D007093:Imidazoles / Q000737:chemistry; D052578:Ionic Liquids / Q000737:chemistry*; D013501:Surface-Active Agents / Q000737:chemistry; D003313:Zea mays / Q000737:chemistry* / Q000235:genetics
D002021:Buffers; D002593:Cetrimonium Compounds; D018744:DNA, Plant; D007093:Imidazoles; D052578:Ionic Liquids; D013501:Surface-Active Agents; C518739:1-ethyl-3-methylimidazolium; D000077286:Cetrimonium