Development of a CTAB buffer-based automated gDNA extraction method for the surveillance of GMO in seed

Seed imported into the EU from countries growing genetically modified (gm) plants may contain traces of these gm crops. As a result of the zero tolerance policy of the EU, these products must be removed from the market. Along with the amount of biotech crops produced worldwide, the work load for see...

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Bibliographic Details
Published in:European food research & technology 2013-04, Vol.236 (4), p.599-606
Main Authors: Guertler, Patrick, Harwardt, Andrea, Eichelinger, Adelina, Muschler, Paul, Goerlich, Ottmar, Busch, Ulrich
Format: Article
Language:English
Subjects:
Agriculture
Analytical Chemistry
Automation
Biotechnology
Chemistry
Chemistry and Materials Science
Corn
Crops
Deoxyribonucleic acid
DNA
Food safety
Food Science
Forestry
Genetically altered foods
Genetically modified organisms
Methods
Original Paper
Polymerase chain reaction
Regulation
Seeds
Soybeans
Studies
Surveillance
Workloads
Zero tolerance
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Summary:Seed imported into the EU from countries growing genetically modified (gm) plants may contain traces of these gm crops. As a result of the zero tolerance policy of the EU, these products must be removed from the market. Along with the amount of biotech crops produced worldwide, the work load for seed surveillance authorities increases. Since the commonly used CTAB buffer-based extraction methods are manual and laborious, a large part of the work load is caused by DNA extraction. In order to reduce labour input and accelerate the DNA analysis workflow, we developed an automated CTAB buffer-based DNA isolation method for seed. Several isolation and chemistry parameters were altered to combine a thorough cell lysis, removal of inhibitors and a highly efficient binding of gDNA to paramagnetic beads. This optimised procedure was compared with manual CTAB buffer-based and Wizard-based DNA extraction methods for maize, soya bean and rapeseed. Automated DNA extraction was faster, less laborious and resulted, on average, in higher DNA yield and purity. The applicability of our method was successfully proven with in-house routine samples.
ISSN:1438-2377
1438-2385
DOI:10.1007/s00217-013-1916-y