BIO NEWS. Food Diagnostics Special. Genomic DNA from food and feed for GMO detection / quantification and animal species differentiation

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1 BIO NEWS Food Diagnostics Special Genomic DNA from food and feed for GMO detection / quantification and animal species differentiation

2 NucleoSpin Food NucleoSpin Food Purification of genomic DNA from food and feed for GMO detection/quantification and animal species differentiation FAST and EASY silica membrane technology HIGH-QUALITY DNA complete removal of PCR inhibitors a variety of complex matrices can be processed DNA ready-to-use for real-time PCR FEATURES sample size: up to 00 mg typical yield: 0. 0 µg DNA elution volume: 00 µl working procedure lysis load lysate st wash APPLICATIONS processed food soya (milk and flour) chocolate cereals meat animal feed nd wash dry elute Ordering information Product Preps Specification Cat.No. NucleoSpin Food 0 0 NucleoSpin columns, collecting tubes ml, buffers CF, C, C, CQW, C, CE, proteinase K 0 0 NucleoSpin columns, collecting tubes ml, buffers CF, C, C, CQW, C, CE, proteinase K 0 0 NucleoSpin columns, collecting tubes ml, buffers CF, C, C, CQW, C, CE, proteinase K Lysis buffer CF litre 70 96

3 NucleoSpin Food NucleoSpin Food Applications GMO quantification in cereals and chocolate A B C D M 6 7 M: Marker : Biscuits : Soya milk : Chocolate : Muesli : Lecithin 6 : Chocolate coating 7 : Soya flour Analysis of Roundup-Ready Soya with LightCycler technology Genomic DNA was purified with NucleoSpin Food from cereals (red) and chocolate (green). DNA content was quantified by amplification with soya lectin-specific primers (Fig. A). Blue curves represent a dilution series of soya standard DNA. Fig. B shows the second amplification of the purified DNA with GMO-specific primers and a dilution series of GMO-standard DNA for the quantification of GMO DNA content. Fig. C: calibration curve for quantification of Roundup-Ready soya. Fig. D: corresponding melting curves of GMO-specific amplification. Soya lectin specific PCR Indicated samples were purified with NucleoSpin Food. Diluted aliquots of the eluates were amplified with lectinspecific primers resulting in a 00 bp fragment. Even from complex and processed matrices like chocolate amplificable DNA could be purified. : Cereals : Chocolate,, : Standard samples (dilution series)

4 NucleoSpin Food Applications Bovine DNA in animal feed M,:,:,6: 7,8: 9,0: M: Feed without animal meal 0. % animal meal 0. % animal meal.0 % animal meal.0 % animal meal Marker Detection of bovine DNA in animal feed, spiked with bovine meal Cattle feed (9 % corn gluten, 7 % brewery residues, % citrus pulp, % barley, 9 % soya meal & % canola meal) was spiked with defined quantities of bovine meal, which had been heat treated at C and. bar over 0 min. DNA was extracted with NucleoSpin Food and amplified with the DNAnimal BOSIdent* kit from GeneScan for specific detection of bovine DNA. The system detects bovine meal at concentrations above 0. % (occasionally down to 0. %). The amplicon is derived from mitochondrial DNA and has a length of only 7 bp. Together with the highly efficient NucleoSpin Food extraction system these characteristics allow the detection of bovine DNA at high sensitivity and specificity even in highly processed food and feed like heattreated animal meal. *BOSIdent (catalogue Nr. 00) is a product of GeneScan Europe AG ( Data kindly provided by GeneScan Europe AG Detection of beef components in food M : : : : : 6: 7: 8: M: Bovine steak, unprocessed Roast beef Sausage without beef Corned beef Beef salami Veal Turkey Pâté (Duck) Marker Beef detection in sausage products DNA preparation was done according to the NucleoSpin Food standard protocol. Aliquots of the 00 µl eluates were amplified with primers and components of a commercial kit (Alcum, Germany). Bovine DNA could be detected in several products, even in strongly processed samples. Sample 8 was declared to be prepared from duck meat only, but clearly showed presence of beef. Samples and 7 did not contain detectable amounts of bovine DNA. Data kindly provided by GEN-IAL, Germany

5 NucleoSpin Food NucleoSpin Food GMO, Food and Feed Analysis with NucleoSpin Food NucleoSpin isolation technology from MACHEREY-NAGEL and GMO experience from GEN-IAL were combined to provide an optimal lysis and purification system for nearly all types of food samples. Resulting eluates are ready-to-use for all types of subsequent detection methods, especially real time and basic PCR technology. GEN-IAL is an established startup company which offers contract research and molecular testing services in food and feed stuff. Special terms of interest are the development and standardisation of detection methods for GMOs and animal and microbial species identification and differentiation. GEN-IAL is the winner of the innovation award 000 of NRW/Germany. The prize was awarded for the First-Beer-Kit for detection of main beer spoilage bacteria. GEN-IAL Institute for Applied Laboratory Analysis Mülheimerstrasse Tor Gebäude 9 D-80 Troisdorf Germany Features High-quality DNA bp 00 bp NucleoSpin Food purified DNA is ideally suitable for amplification DNA from soya meal was purified with NucleoSpin Food and used for PCR amplification with lectin- (lanes,) and plant- (lanes,) specific primer-pairs. M... suitable for real-time PCR A Chocolate Biscuits Muesli DNA analysis with LightCycler technology NucleoSpin Food purified DNA from three matrices was amplified with lectin-specific primers and SYBR-Green detection mode in a LightCycler system (A). Specific amplification was verified by a melting point analysis (B). B Chocolate Biscuits Muesli

6 Purification of Genomic DNA Purification of Genomic DNA Bio FD e/0/0/.00 DP Printed in Germany Overview NucleoSpin Kits for purification of Genomic DNA Tissue, cells and forensic samples NucleoSpin Tissue kits Product format feature typical applications NucleoSpin Tissue mini spin columns high capacity, genotyping (HLA), high DNA recovery forensics NucleoSpin Multi-8 Tissue 8-well strips medium-throughput genotyping, PCR NucleoSpin Multi-96 Tissue 96-well plates high-throughput genotyping, PCR Plant and fungi NucleoSpin Plant kits Product format feature typical applications NucleoSpin Plant mini spin columns high capacity, genotyping, PCR, high DNA recovery southern blotting NucleoSpin Plant L midi spin columns high capacity, genotyping, PCR, high DNA recovery southern blotting NucleoSpin Plant XL maxi spin columns high capacity, genotyping, PCR, high DNA recovery southern blotting NucleoSpin Multi-8 Plant 8-well strips medium-throughput genotyping, PCR, southern blotting NucleoSpin Multi-96 Plant 96-well plates high-throughput genotyping, PCR, southern blotting Food NucleoSpin Food mini spin columns high sensitivity, genotyping, PCR high DNA recovery If you have any questions, please contact our technical service: Tel.: Fax: tech-bio@macherey-nagel.de Please visit our internet shop: MACHEREY-NAGEL sales@macherey-nagel.de MACHEREY-NAGEL GmbH & Co. KG. P.O. Box 0. D- Düren. Germany. Tel. (0 ) Fax (0 ) Switzerland: MACHEREY-NAGEL AG. P.O. Box. CH-70 Oensingen. Tel. (0 6) Fax (0 6) 88 0 France: MACHEREY-NAGEL S.àr.l.., rue Gutenberg. B.P.. F-677 Hoerdt. Tel Fax USA: MACHEREY-NAGEL Inc. 6 South Third St., Suite 0 Easton, PA 80 Tel Fax TÜV CERT DIN EN ISO 900 Certificate: