Anion-exchange chromatography using CIM® DEAE disks as a method of choice for DNA isolation from lecithin

Authors

  • Špela Höfferle
  • Ana Kramar
  • Franc Smrekar
  • Dejan Štebih
  • Jana Vojvoda
  • Jana Žel
  • Peter Raspor

DOI:

https://doi.org/10.14720/abs.50.1.14438

Keywords:

DNA isolation, detection of GMO, soybean lecithin, chromatography, monoliths, CIM, CTAB, PCR, real-time PCR

Abstract

The most important prerequisite for the application of PCR-based methods, among them the detection and quantifi cation of genetically modifi ed organisms (GMOs) is the ability to extract significant amounts of DNA of adequate quality from the sample under investigation. The sample of interest in our work was soybean lecithin with expected low DNA content. The aim of this study was to set up a fast and effective HPLC (High Performance Liquid Chromatography) method using CIM® (Convective Interaction Media, BIA Separations d.o.o., Ljubljana, Slovenia) DEAE (DiEthylAminoEthyl) anion-exchange disk monolithic columns (disks) for the isolation of DNA from soybean lecithin samples. As the reference isolation procedure we used CTAB (CethylTrimethylAmmonium Bromide) method, which is widely used in GMO detection. It was demonstrated, that CIM® DEAE disks allow effi cient isolation of DNA from soybean lecithin. Furthermore, in comparison with the CTAB method, the method was less time-consuming and reduced the use of some aggressive chemicals. The quality of isolated DNA was tested with spectrophotometric analysis, agarose gel electrophoresis and by amplification of soybean specifi c lectin gene with qualitative and real-time PCR. The isolated soybean DNA was of adequate quantity and quality for PCR analysis, even though mostly degraded, present in small amounts and contaminated with some impurities, among them potential PCR inhibitors. The study expanded the applicability of monolithic columns in the isolation of biomolecules from highly processed food materials and their potential use for nucleic acids detection.

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Published

01.07.2007

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Original Research Paper

How to Cite

Höfferle, Špela, Kramar, A., Smrekar, F., Štebih, D., Vojvoda, J., Žel, J., & Raspor, P. (2007). Anion-exchange chromatography using CIM® DEAE disks as a method of choice for DNA isolation from lecithin. Acta Biologica Slovenica, 50(1), 19-29. https://doi.org/10.14720/abs.50.1.14438

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