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A simple capillary gel electrophoresis approach for efficient and reproducible DNA separations. Analysis of genetically modified soy and maize
Author(s) -
Sánchez Laura,
González Ramón,
Crego Antonio L.,
Cifuentes Alejandro
Publication year - 2007
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200600195
Subject(s) - capillary electrophoresis , chromatography , chemistry , genetically modified maize , electrophoresis , genetically modified organism , capillary action , hydroxyethyl cellulose , cellulose , dna , biochemistry , materials science , genetically modified crops , transgene , composite material , gene
It is generally assumed that in order to achieve suitable separations of DNA fragments, capillary gel electrophoresis (CGE)‐coated capillaries should be used. In this work, a new method is presented that allows to obtain reproducible CGE separations of DNA fragments using bare fused‐silica capillaries without any previous coating step. The proposed method only requires: (i) a capillary washing with 0.1 M hydrochloric acid between injections and (ii) a running buffer composed of Tris‐phosphate‐ethylenediamine tetraacetic acid (EDTA) and 4.5% of 2‐hydroxyethyl cellulose (HEC) as sieving polymer. The use of this new CGE procedure gives highly resolved and reproducible separations of DNA fragments ranging from 50 to 750 bp. The separation of these DNA fragments is accomplished in less than 30 min with efficiencies up to 1.7×10 6 plates/m. Reproducibility values of migration times (given as %RSD) for the analyzed DNA fragments are better than 1.0% ( n = 4) for the same day, 2.2% ( n = 16) for four different days, and 2.3% ( n = 16) for four different capillaries. The usefulness of this separation method is demonstrated by detecting genetically modified maize and genetically modified soy after DNA amplification by PCR. This new CGE procedure together with LIF as detector provides sensitive analysis of 0.9% of Bt11 maize, Mon810 maize, and Roundup Ready soy in flours with S/N up to 542. These results demonstrate the usefulness of this procedure to fulfill the European regulation on detection of genetically modified organisms in foods.