Detection of processed genetically modified food using CIM monolithic columns for DNA isolation.
The availability of sufficient quantities of DNA of adequate quality is crucial in polymerase chain reaction (PCR)-based methods for genetically modified food detection. In this work, the suitability of anion-exchange CIM (Convective Interaction Media; BIA Separations, Ljubljana, Slovenia) monolithic columns for isolation of DNA from food was studied. Maize and its derivates corn meal and thermally pretreated corn meal were chosen as model food. Two commercially available CIM disk columns were tested: DEAE (diethylaminoethyl) and QA (quaternary amine). Preliminary separations were performed with standard solution of salmon DNA at different pH values and different NaCl concentrations in mobile phase. DEAE groups and pH 8 were chosen for further isolations of DNA from a complex matrix-food extract. The quality and quantity of isolated DNA were tested on agarose gel electrophoresis, with UV-scanning spectrophotometry, and by amplification with real-time PCR. DNA isolated in this way was of suitable quality for further PCR analyses. The described method is also applicable for DNA isolation from processed foods with decreased DNA content. Furthermore, it is more effective and less time-consuming in comparison with the existing proposed methods for isolation of DNA from plant-derived foods.
Jerman
Sergej
S
Department of Food Science and Technology, Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, S-1000 Ljubljana, Slovenia.
Podgornik
Ales
A
Cankar
Katarina
K
Cadet
Neza
N
Skrt
Mihaela
M
Zel
Jana
J
Raspor
Peter
P
D002852:Chromatography, Ion Exchange / Q000295:instrumentation*; D004247:DNA / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D030841:Food, Genetically Modified; D016133:Polymerase Chain Reaction; D013056:Spectrophotometry, Ultraviolet
D004247:DNA