Guidelines for validation of DNA extraction methods applied in subsequent PCR analysis of food and feed products for the presence of genetically modified material

2014 ◽  
Vol 9 (2) ◽  
pp. 183-190 ◽  
Author(s):  
Hans-Ulrich Waiblinger ◽  
Lutz Grohmann
Keyword(s):  
Dna Extraction ◽  
Genetically Modified ◽  
Extraction Methods ◽  
Pcr Analysis ◽  
Food And Feed ◽  
Dna Extraction Methods

scholarly journals Determining the optimal method for DNA isolation from fruit jams

2018 ◽  
Vol 36 (No. 2) ◽  
pp. 126-132
Author(s):  
Sovová Tereza ◽  
Křížová Barbora ◽  
Ovesná Jaroslava
Keyword(s):  
Dna Extraction ◽  
Dna Isolation ◽  
Extraction Methods ◽  
Food Samples ◽  
Pcr Analysis ◽  
Uv Spectrophotometry ◽  
Pcr Assay ◽  
Optimal Method ◽  
Ctab Method ◽  
Dna Extraction Methods

DNA extraction is a crucial step in PCR analysis especially when analysing food samples that can be degraded and can potentially contain PCR-inhibiting substances. In this study, we compared the suitability of three DNA extraction methods – two kits: DNeasy<sup>®</sup> Plant Mini Kit and NucleoSpin<sup>®</sup> Food, and the CTAB method – for DNA extraction from commercial fruit jams. Fourteen jams with different contents of fruit, sugar and other additives were extracted in triplicate using the above-mentioned methods directly and after a washing step. The concentration and optical density were analysed using UV spectrophotometry and the amplifiability of the obtained DNA was evaluated using a PCR assay targeting a sequence coding for chloroplast tRNA-Leu. Samples isolated using the NucleoSpin<sup>®</sup> Food kit contained non-amplifiable DNA in eight cases, and samples isolated using the CTAB method could not be quantified. The DNeasy<sup>®</sup> Plant Mini Kit thus proved to be the most suitable method, since well-amplifiable DNA was obtained for all the analysed samples.

DNA extraction methods for detecting genetically modified foods: A comparative study

2011 ◽  
Vol 126 (4) ◽  
pp. 1883-1889 ◽  
Author(s):  
Rafaat M. Elsanhoty ◽  
Mohamed Fawzy Ramadan ◽  
Klaus Dieter Jany
Keyword(s):  
Comparative Study ◽  
Dna Extraction ◽  
Genetically Modified ◽  
Extraction Methods ◽  
Genetically Modified Foods ◽  
Dna Extraction Methods

Evaluation of DNA extraction methods and dilution treatment for detection and quantification of Acanthamoeba in water and biofilm by real-time PCR

2010 ◽  
Vol 62 (9) ◽  
pp. 2141-2149 ◽  
Author(s):  
Ching-Wen Chang ◽  
Ying-Chieh Wu
Keyword(s):  
Real Time ◽  
Dna Extraction ◽  
Real Time Pcr ◽  
Pathogenic Bacteria ◽  
Acanthamoeba Castellanii ◽  
Extraction Methods ◽  
Cell Number ◽  
Pcr Analysis ◽  
Human Pathogens ◽  
Dna Extraction Methods

Acanthamoeba, human pathogens and natural hosts of pathogenic bacteria, may be accurately detected and quantified by real-time PCR if Acanthamoeba DNA are properly extracted and PCR inhibitors are effectively eliminated. However, the optimization of DNA extraction methods has not been reported for Acanthamoeba. This study compared the effectiveness of two DNA extraction/purification methods (FastDNA® Spin Kit for soil and Wizard® SV genomic DNA Purification System) by using trophozoites and cysts of Acanthamoeba castellanii and water and biofilm samples of cooling towers. DNA of A. castellanii extracted with the FastDNA® Kit and quantified by TaqMan PCR resulted in a lower variation (CV of Ct &lt; 3%), greater linearity (R2=0.99), and higher slopes (1.177–1.187 log fg DNA/log cell number) as compared to that by the Wizard® Kit. For field testing, the number of Acanthamoeba-positive samples and the Acanthamoeba DNA quantity were both greater with the FastDNA® Kit than with the Wizard® Kit (P=0.016 and &lt;0.0001, respectively). Beneficial effects with dilutions of extracted DNA were also revealed with the FastDNA® Kit (P=0.0003). In conclusion, DNA extraction by the FastDNA® Kit coupled with dilution of extracted DNA and PCR analysis are recommended for detecting and quantifying environmental Acanthamoeba.

Evaluation of DNA extraction methods for Bacillus anthracis spores spiked to food and feed matrices at biosafety level 3 conditions

2011 ◽  
Vol 150 (2-3) ◽  
pp. 122-127 ◽  
Author(s):  
Peter R. Wielinga ◽  
Lianne de Heer ◽  
Astrid de Groot ◽  
Raditijo A. Hamidjaja ◽  
Geert Bruggeman ◽  
...  
Keyword(s):  
Bacillus Anthracis ◽  
Dna Extraction ◽  
Extraction Methods ◽  
Bacillus Anthracis Spores ◽  
Food And Feed ◽  
Level 3 ◽  
Dna Extraction Methods ◽  
Biosafety Level
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