Certification of Reference Materials of Soya Seed Powder with different Mass Fractions of genetically modified roundup ready® Soya, Certified Reference Materials ERM®-BF410k (ERM®-BF410ak, ERM®-BF410bk, ERM®-BF410dk, ERM®-BF410gk)

This report descirbes the processing and certification of four soya seed powder Certified Reference Materials (CRMs) containing different mass fractions of genetically modified (GM) GTS 40-30-2 (Roundup Ready) soya (ERM-BF410ak, bk, dk, gk), according to the principles of ISO Guide 34. The certification values were confirmed by event-specific real-time PCR as independent verification method. The CRMs are intended to be used for quality control or calibration of methods for the quantification of the GTS 40-30-2 soya mass fraction in food and feed. The four CRMs have been accepted as European Reference Material (ERM®) after peer evaluation by the partners of the European Reference Material consortium.JRC.D.2-Reference material

Certification of Plasmid DNA containing NK603 Maize DNA Fragments - Certified Reference Material ERM®-AD415

This report describes the preparation, characterisation, stability and suitability studies of the certified reference material (CRM) ERM-AD415 which contains a plasmid (pIRMM-0086) carrying a defined deoxyribonucleic acid (DNA) fragment specific for a genetic modification present in Zea Mays L. line NK603 as well as defined DNA fragments specific for the Zea mays taxon, of which one has been certified. The CRM was processed in 2009 and certified in 2010 by the European Commission, Joint Research Centre, Institute for Reference Materials and Measurements (IRMM), Geel, BEJRC.DG.D.2-Reference material

Certification of Reference Materials of Cotton Seed Powder with Different Mass Fractions of the Cotton Event GHB119 - Certified Reference Materials ERM®BF428 (ERM®-BF428a, ERM®-BF428b, ERM®-BF428c)

This report describes the processing and certification of three cotton seed powder Certified Reference Materials (CRMs) containing different mass fractions of genetically modified (GM) GHB119 cotton (ERM-BF428a, b, c). The materials were processed and certified in 2008/2009 by the European Commission, Joint Research Centre, Institute for Reference Materials and Measurements (IRMM, Geel, Belgium), according to the principles of ISO Guide 34. The three CRMs have been accepted as European Reference Materials (ERM) after peer evaluation by the partners of the European Materiasl consortium.JRC.D.2-Reference material

Certification of Reference Materials of Soya Seed Powder with different Mass Fractions of Genetically Modified 305423 Soya, Certified Reference Materials ERM-BF426 (ERM-BF426a, ERM-BF426b, ERM-BF426c, ERM-BF426d)

This report describes the processing and certification of four soya seed powder Certified Reference Materials (CRMs) containing different mass fractions of genetically modified (GM) 305423 soya (ERM-BF426a, b, c, d). The materials were processed and certified in 2007 by the European Commission, Directorate General Joint Research Centre, Institute for Reference Materials and Measurements (IRMM, Geel, Belgium), according to the principles of ISO Guide 34. The CRMs are intended to be used for quality control or calibration of methods for the quantification of the 305423 soya mass fraction in food and feed. The CRMs are available in glass bottles containing 1 g of dried soya powder closed under argon atmosphere. The minimum amount of sample to be used per analysis is 200 mg. The four CRMs have been accepted as European Reference Material (ERM) after peer evaluation by the partners of the European Reference Materials consortium.JRC.D.2-Reference material

Certification of Reference Materials of Maize Seed Powder containing Genetically Modified MON 810 Maize: Certified Reference Materials ERM®-BF413k (ERM®-BF413ak, ERM®-BF413ck, ERM®-BF413ek, ERM®-BF413gk)

The report describes the processing and certification of four maize seed powder Certified Reference Materials (CRMs) containing different mass fractions of genetically modified (GM) MON 810 maize (ERM-BF413ak, ck, ek, gk). The materials were processed and certified in 2007/2008 by the European Commission, Joint Research Centre, Institute for Reference Materials and Measurements IRMM, Geel, Belgium), according to the principles of ISO Guide 34. The four CRMs belonging to the ERM-BF413k series replace the earlier produced series ERM-BF413 which is exhausted. The four CRMs have been accepted as European Reference Material (ERM®) after peer evaluation by the partners of the European Reference Materials consortium.JRC.D.2-Reference material

Oligonucleotide batch quality has a limited impact on quantitative real-time PCR

Quantitative real-time PCR (qPCR) is currently the method of choice for sequence specific quantification of DNA. The design of oligonucleotide primers and probes is essential. Oligonucleotides are available not only from different manufacturers but also in different purity grades. Three event-specific qPCR methods for the quantification of genetically modified organisms (GMOs) were chosen, namely for maize NK603, maize MIR604 and soya 356043 to assess a possible impact of the choice of manufacturer and purity grade of oligonucleotides on qPCR results. Coefficients of determination (R2), slopes and y-intercepts obtained from calibration curves using oligonucleotides of different suppliers and purity grades were evaluated statistically and compared. The same oligonucleotides were used to quantify the DNA content of GMO in Certified Reference Materials (CRMs) ERM-BF415f (maize NK603), ERM-BF423d (maize MIR604) and ERM-BF425d (soya 356043) by qPCR. All calibration curves for all events and all suppliers and purity grades were highly linear. The lowest R2 observed in this study was 0.997. No considerable differences were observed for mean R2 from calibration curves generated with oligonucleotides from different suppliers and purity grades. For mean slopes and mean y-intercepts of calibration curves group distribution patterns per PCR target were observed. However, for all PCR targets no tendency in performance between suppliers and purity grades could be identified. A statistical comparison of slopes was performed testing the equivalence of all analysed calibration curves. Though slopes were significantly different for some of the investigated PCR targets, no effect of oligonucleotide supplier or purity grade could be shown. Also, the estimated GMO content for the three CRMs ERM-BF415f, ERM-BF423d and ERM-BF425d using different sources and purity grades of oligonucleotides were in agreement with each other with relative standard deviations below 4 %. With this study we demonstrate that the choice of oligonucleotide manufacturer and purity grade has little impact on qPCR measurement results. Consequently, we suggest that no additional uncertainty component has to be accounted for in the uncertainty budget of qPCR measurement results. However, oligonucleotide quality can have an effect on qPCR calibration especially in qPCR methods that tend to be less robust.JRC.D.2-Standards for Innovation and sustainable Developmen

Certification of a Maize 98140 Reference Material for its DNA Copy Number Ratio - Certified Reference Materials ERM®-BF427c

This report describes the certification of the Certified Reference Material (CRM) ERM-BF427c for its DNA copy number ratio. This CRM is a powder processed from seeds of genetically modified 98140 maize and conventional maize. ERM-BF427c is part of a set of maize CRMs containing different mass fractions of GM maize 98140. The CRM was processed and originally certified for its mass fraction by the European Commission, Joint Research Centre, Institute for Reference Materials and Measurements (IRMM), Geel, BEJRC.DG.D.2-Reference material

Certification of a Soya 356043 Reference Material for its DNA Copy Number Ratio - Certified Reference Materials ERM®-BF425c

ERM-BF425c is part of a set of soya CRMs containing different mass fractions of GM soya 356043. The CRM was processed and originally certified for its soya 356043 mass fraction by the European Commission, Joint Research Centre, Institute for Reference Materials and Measurements (IRMM), Geel, BE. European Commission Recommendation (EC) No 787/2004 advises to express the content of GM food and feed products as the percentage of GM DNA copy numbers in relation to target taxon-specific DNA copy numbers calculated in terms of haploid genomes, further referred to as the DNA copy number ratio. An interlaboratory comparison was conducted in 2008 to certify the existing CRM for its DNA copy number ratio. The CRM is intended for quality control of measurements of DNA copy number ratios of soya event 356043 in GM food and feed. Hereby, the plasmid DNA (pDNA) ERM®-AD425 should be used for calibration together with the soya 356043 event-specific real-time Polymerase Chain Reaction method validated by the European Union Reference Laboratory for GM Food and Feed (EURL-GMFF) (available via: http://gmo-crl.jrc.ec.europa.eu/statusofdoss.htm). ERM-AD425 contains a 99 bp fragment of the 5' plant/insert junction specific for the soya event 356043. Additionally, the plasmid carries a 243 bp fragment of the species-specific lectin gene le1, specific for the soya taxon. The CRM ERM-BF425c is available in glass bottles containing 1 g of soya seed powder, closed under argon atmosphere and is certified for its DNA copy number ratio. The minimum amount of sample to be used is 200 mg.JRC.DG.D.2-Reference material

Certification of Reference Materials of Maize Seed Powder with different Mass Fractions of Genetically Modified 98140 Maize, ERM-BF427 (ERM-BF427a, ERM-BF427b, ERM-BF427c, ERM-BF427d)

This report describes the processing and certification of four maize seed powder Certification Reference Materials (CRMs) containing different mass fractions of genetically modified (GM) DP-ø9814ø-6 maize (ERM-BF427a, b, c, d), according to the principles of ISO Guide 34. The certification values were confirmed by event-specific real-time PCR as independent verification method. The CRMs are intended to be used for quality control of calibration of methods for the quantification of the DP-ø9814ø-6 maize mass fraction in food and feed. The four CRMs have been accepted as European Reference Materials (ERM) after peer evaluation by the partners of the European Reference Materials consortium.JRC.D.2-Reference material