Test-systems for GMO detection and identification

test-systems are used to perform one stage detection and identification of GMO of plant origin with polymerase chain reaction in real time mode in matrix format;
78 test systems include primers, probes and plasmids (positive control) for identification of 31 transformation events (including all lines approved in this or that form in the Russian Federation), 17 sense sequences, 13 regulatory sequences (promoters, terminators), 4 selective marker genes and 13 species of main agricultural crops undergoing genetic engineering to reveal species identity of a test sample of undefined composition;
test systems arranged on a matrix permit one to identify simultaneously 31 transformational events of agricultural GM crops in one experiment (within 35-40 minutes) and at the same time detect by genetic elements majority of GM crop plants authorized worldwide (maize: ~ 91%, soybean: ~ 92%, rapeseed: ~ 93% ...);
test systems have been validated using reference materials provided by the FGBUN "Federal Research Center for Nutrition and Biotechnology" as the part of the regulatory activites - development of Methodological Guidelines "Detection and Identification of GMO of Plant Origin with Polymerase Chain Reaction in Matrix Format" (MUK 4.2.3390-16), as well as standard reference materials produced by ERM JRC EU, American Oil Chemists' Society and Sigma-Aldrich;
main test-systems used are validated by ring-testing, reported in the EU Compendium of Methods, and confirmed by member organizations of the European Network of GMO Laboratories (ENGL)

Lines

Object	Target	Oligonucleotides	Refernce
Maize<br><b>176</b>	3'-border region between insert and maize genome<br>Amplicon length: 82 b.p.	PRF: GGCCGTGAACGAGCTGTT<br>PRR: GGGAAGAAGCCTACATGTTTTCTAA<br>PR: FAM-AGCAACCAGATCGGCCGACACC-BHQ1	Savini C, Bonfini L, Querci M, Mazzara M, Cordeil S, Van den Eede G. Event-specific Method for the Quantitation of Maize Line Bt176 Using Real-Time PCR - Validation Report and Protocol. 2011.
Maize<br><b>3272</b>	5'-border region between insert and maize genome<br>Amplicon length: 95 b.p.	PRF: TCATCAGACCAGATTCTCTTTTATGG<br>PRR: CGTTTCCCGCCTTCAGTTTA<br>PR: FAM-ACTGCTGACGCGGCCAAACACTG-BHQ1	Charles Delobel C, Foti N, Mazzara M, Van den Eede G. Event-specific Method for the Quantitation of Maize Event 3272 Using Real-Time PCR - Validation Report and Protocol. 2008. JRC48922
Maize<br><b>5307</b>	3'-border region between insert and maize genome<br>Amplicon length: 107 b.p.	PRF: CATGGCCGTATCCGCAATGTG<br>PRR: TGCACCCTTTGCCAGTGG<br>PR: FAM-ACCACAATATACCCTCTTCCCTGGGCCAG-BHQ1	Hart H. Event 5307 Maize. Real-time, Event-specific Polymerase Chain Reaction Method. Annex B. 2011
Maize<br><b>Bt11</b>	3'-border region between insert and maize genome<br>Amplicon length: 70 b.p.	PRF: GCGGAACCCCTATTTGTTTA<br>PRR: TCCAAGAATCCCTCCATGAG<br>PR: ROX-AAATACATTCAAATATGTATCCGCTCA-BHQ2	Mazzara M, Puumalaainen J, Van den Eede G. Validation of the GMO Specific Detection Method Developed by NVI/NRA for Bt11 in Sweet Corn Maize - Validation Report and Protocol. 2005. JRC32111
Maize<br><b>GA21</b>	5'-border region between insert and maize genome<br>Amplicon length: 112 b.p.	PRF: CTTATCGTTATGCTATTTGCAACTTTAGA<br>PRR: TGGCTCGCGATCCTCCT<br>PR: ROX-CATATACTAACTCATATCTCTTTCTCAACAGCAGGTGGGT-BHQ2	Paoletti C, Mazzara M, Puumalaainen J, Rasulo D, Van den Eede G. Event-specific Method for the Quantitation of Maize Line GA21 Using Real-Time PCR - Validation Report and Protocol. 2005. JRC32087
Maize<br><b>LY038</b>	5'-border region between insert and maize genome<br>Amplicon length: 112 b.p.	PRF: TGGGTTCAGTCTGCGAATGTT<br>PRR: AGGAATTCGATATCAAGCTTATCGA<br>PR: FAM-CGAGCGGAGTTTATGGGTCGACGG-BHQ1	Charles Delobel C, Grazioli E, Larcher S, Mazzara M, Van den Eede G. Event-specific Method for the Quantification of Maize Line LY038 Using Real-time PCR - Validation Report and Protocol. EUR 23647 EN. Luxembourg (Luxembourg): Publications Office of the European Union, 2008. JRC48919
Maize<br><b>MIR162</b>	3'-border region between insert and maize genome<br>Amplicon length: 92 b.p.	PRF: GCGCGGTGTCATCTATGTTACTAG<br>PRR: TGCCTTATCTGTTGCCTTCAGA<br>PR: FAM-TCTAGACAATTCAGTACATTAAAAACGTCCGCCA-BHQ1	Charles Delobel C, Mazzara M, Bevilacqua A, Van den Eede G. Event-specific Method for the Quantitation of Maize MIR162 by Real-time PCR. Validation Report and Protocol. 2011. JRC68155
Maize<br><b>MIR604</b>	5'-border region between insert and maize genome<br>Amplicon length: 76 b.p.	PRF: GCGCACGCAATTCAACAG<br>PRR: GGTCATAACGTGACTCCCTTAATTCT<br>PR: ROX-AGGCGGGAAACGACAATCTGATCATG-BHQ2	Mazzara M, Munaro B, Foti N, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Maize Line MIR604 Using Real-Time PCR - Validation Report and Protocol. 2007. JRC37490
Maize<br><b>MON810<b>	5'-border region between insert and maize genome<br>Amplicon length: 92 b.p.	PRF: TCGAAGGACGAAGGACTCTAACGT<br>PRR: GCCACCTTCCTTTTCCACTATCTT<br>PR: FAM-AACATCCTTTGCCATTGCCCAGC-BHQ1	ISO 21569:2005: Foodstuffs-Methods of analysis for the detection of genetically modified organisms and derived products-Qualitative nucleic acid based methods
Maize<br><b>MON863</b>	5'-border region between insert and maize genome<br>Amplicon length: 84 b.p.	PRF: TGTTACGGCCTAAATGCTGAACT<br>PRR: GTAGGATCGGAAAGCTTGGTAC<br>PR: ROX-TGAACACCCATCCGAACAAGTAGGGTCA-BHQ2	Mazzara M, Foti N, Price S, Paoletti C, Van den Eede G. Event-specific Method for the Quantitation of Maize Line MON 863 Using Real-Time PCR - Validation Report and Protocol. 2005. JRC32105
Maize<br><b>MON88017</b>	3'-border region between insert and maize genome<br>Amplicon length: 94 b.p.	PRF: GAGCAGGACCTGCAGAAGCT<br>PRR: TCCGGAGTTGACCATCCA<br>PR: ROX-TCCCGCCTTCAGTTTAAACAGAGTCGGGT-BHQ2	Charles Delobel C, Foti N, Grazioli E, Mazzara M, Van den Eede G. Event-specific Method for the Quantitation of Maize Line MON 88017 Using Real-Time PCR - Validation Report and Protocol. 2008. JRC48920
Maize<br><b>MON89034</b>	3'-border region between insert and maize genome<br>Amplicon length: 77 b.p.	PRF: TTCTCCATATTGACCATCATACTCATT<br>PRR: CGGTATCTATAATACCGTGGTTTTTAAA<br>PR: FAM–ATCCCCGGAAATTATGTT–MGBNFQ	Savini C, Bogni A, Grazioli E, Munaro B, Mazzara M, Van den Eede G. Event-specific Method for the Quantitation of Maize Line MON 89034 Using Real-Time PCR - Validation Report and Protocol. 2008. JRC48921
Maize<br><b>MZIR098</b>	5'-border region between insert and maize genome<br>Amplicon length: 73 b.p.	PRF: ATCTCAGACACCAAACCGAGATC<br>PRR: ACACCGTTAGGCTAGTGCCAGT<br>PR: FAM-CAAGTGACAGCGAACGGAGCTGGTTT-BHQ1	European Union Reference Laboratory for GM Food and Feed (EURL GMFF), Joint Research Centre (JRC), European Commission. Event-specific Method for the Quantification of Maize MZIR098 Real-time PCR - Validation Report. 2018. JRC 114043. Online Publication
Maize<br><b>MZHG0JG</b>	3'-border region between insert and maize genome<br>Amplicon length: 153 b.p.	PRF: TTAGCTAACGGCCAGGATC<br>PRR: CACCGTGACATGCTTAGCA<br>PR: FAM-CGCGTGAGCCTTTAGCAACTAGC-BHQ1	GenBit
Maize<br><b>NK603</b>	3'-border region between insert and maize genome<br>Amplicon length: 108 b.p.	PR: ATGAATGACCTCGAGTAAGCTTGTTAA<br>PRR: AAGAGATAACAGGATCCACTCAAACACT<br>PR: ROX-TGGTACCACGCGACACACTTCCACTC-BHQ2	Mazzara M, Paoletti C, Puumalaainen J, Rasulo D, Van den Eede G. Event-specific Method for the Quantitation of Maize Line NK603 Using Real-Time PCR - Validation Report and Protocol. 2005. JRC32103
Maize<br><b>T25</b>	3'-border region between insert and maize genome<br>Amplicon length: 102 b.p.	PRF: ACAAGCGTGTCGTGCTCCAC<br>PRR: GACATGATACTCCTTCCACCG<br>PR: ROX-TCATTGAGTCGTTCCGCCATTGTCG-BHQ2	Mazzara M, Grazioli E, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Maize Line T25 Using Real-Time PCR v. 1.01 - Validation Report and Validated Method. 2013. JRC84152
Maize<br><b>TC1507</b>	3'-border region between insert and maize genome<br>Amplicon length: 58 b.p.	PRF: TAGTCTTCGGCCAGAATGG<br>PRR: CTTTGCCAAGATCAAGCG<br>PR: FAM-TAACTCAAGGCCCTCACTCCG-BHQ1	Mazzara M. Foti N, Price S, Paoletti C, Van den Eede G. Event-specific Method for the Quantitation of Maize Line TC1507 Using Real-Time PCR - Validation Report and Protocol. 2005. JRC32120
Maize<br><b>DAS-40278-9</b>	5'-border region between insert and maize genome<br>Amplicon length: 98 b.p.	PRF: CACGAACCATTGAGTTACAATC<br>PRR: TGGTTCATTGTATTCTGGCTTTG<br>PR: FAM-CGTAGCTAACCTTCATTGTATTCCG-BHQ1	Savini C, Bogni A, Foti N, Mazzara M, Kreysa J. Event-specific Method for the Quantification of Maize DAS-40278-9 by Real-time PCR - Validation Report and Validated Method - Sampling and DNA Extraction of Maize TC15O7. EUR 25585 EN. Luxembourg (Luxembourg): Publications Office of the European Union, 2012
Rapeseed<br><b>GT73 (RT73)</b>	3'-border region between insert and rapeseed genome<br>Amplicon length: 108 b.p.	PRF: CCATATTGACCATCATACTCATTGCT<br>PRR: GCTTATACGAAGGCAAGAAAAGGA<br>PR: FAM-TTCCCGGACATGAAGATCATCCTCCTT-BHQ1	Mazzara M, Grazioli E, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Oilseed Rape Line RT73 Using Real-Time PCR - Validation Report and Protocol. 2007. JRC37550
Rice<br><b>LLRICE62</b>	3'-border region between insert and rice genome<br>Amplicon length: 88 b.p.	PRF: AGCTGGCGTAATAGCGAAGAGG<br>PRR: TGCTAACGGGTGCATCGTCTA<br>PR: FAM-CGCACCGATTATTTATACTTTTAGTCCACCT-BHQ1	Mazzara M, Grazioli E, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Rice Line LLRICE62 Using Real-Time PCR - Validation Report and Protocol. 2006. JRC34091
Sugar beet<br><b>H7-1</b>	5'-border region between insert and sugar beet genome<br>Amplicon length: 108 b.p.	PRF: TGGGATCTGGGTGGCTCTAACT<br>PRR: AATGCTGCTAAATCCTGAG<br>PR: FAM-AAGGCGGGAAACGACAATCT-BHQ1	Mazzara M, Foti N, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Sugar beet line H7-1 Using Real-Time PCR v. 1.01 - Validation Report and Valideted Method. 2013. JRC84121
Soybean<br><b>A2704-12</b>	region containing inverted 3'- and 5'-sequences of bla gene<br>Amplicon length: 64 b.p.	PRF: GCAAAAAAGCGGTTAGCTCCT<br>PRR: ATTCAGGCTGCGCAACTGTT<br>PR: ROX-GGAAGGGCGATCGGAGGACCG-BHQ2	Mazzara M, Querci M, Delobel C, Moens W, Cordeil S, Van den Eede G. Event-specific Method for the Quantification of Soybean Line A2704-12 Using Real-Time PCR. Protocol. 2007
Soybean<br><b>A5547-127</b>	5'-border region between insert and soybean genome<br>Amplicon length: 71 b.p.	PRF: CTATTTGGTGGCATTTTTCC<br>PRR: TGCGGCCAACTTACTTC<br>PR: FAM-ACAACGATGACGGTATGACATTGCGG-BHQ1	Mazzara M, Querci M, Luque-Perez E, Ermolli M, Cordeil S, Van den Eede G. Event-specific Method for the Quantification of Soybean Event A5547-127 Using Real-Time PCR. Protocol. 2009
Soybean<br><b>BPS CV-127-9</b>	3'-border region between insert and soybean genome<br>Amplicon length: 88 b.p.	PRF: AACAGAAGTTTCCGTTGAGCTTTAAGAC<br>PRR: CATTCGTAGCTCGGATCGTGTAC<br>PR: FAM-TTTGGGGAAGCTGTCCCATGCCC-BHQ1	Mazzara M, Querci M, Savini C, Bonfini L, Cordeil S, Van den Eede G. Event-specific Method for the Quantification of Soybean CV127 Using Real-Time PCR. Protocol. 2011
Soybean<br><b>DP305423-1</b></b>	3'-border region between insert and soybean genome<br>Amplicon length: 93 b.p.	PRF: CGTGTTCTCTTTTTGGCTAGC<br>PRR: GTGACCAATGAATACATAACACAAACTA<br>PR: FAM-TGACACAAATGATTTTCATACAAAAGTCGAGA-BHQ1	Mazzara M, Munaro B, Delobel C, Grazioli E, Savini C, Van den Eede G. Event-specific Method for the Quantification of Soybean Event DP-305423-1 Using Real-Time PCR v. 1.01. Validation Report and Validated Method. 2013
Soybean<br><b>FG72</b>	3'-border region between insert and soybean genome<br>Amplicon length: 70 b.p.	PRF : AGATTTGATCGGGCTGCAGG<br>PRR: GCACGTATTGATGACCGCATTA<br>PR: FAM–AATGTGGTTCATCCGTCTT–MGBNFQ	Savini et al. Event-specific Method for the Quantification of Soybean FG72 Using Real-Time PCR. Validation Report and Protocol. 2012
Soybean<br><b>GTS 40-3-2</b>	5'-border region between insert and soybean genome<br>Amplicon length: 84 b.p.	PRF: TTCATTCAAAATAAGATCATACATACAGGTT<br>PRR: GGCATTTGTAGGAGCCACCTT<br>PR: MGB FAM–CCTTTTCCATTTGGG–MGBNFQ	Mazzara M, Querci M, Moens W, Grazioli E, Cordeil S, Van den Eede G. Event-specific Method for the Quantification of Soybean Line 40-3-2 Using Real-Time PCR. Protocol. 2009.
Soybean<br><b>MON87701</b>	5'-border region between insert and soybean genome<br>Amplicon length: 89 b.p.	PRF: TGGTGATATGAAGATACATGCTTAGCAT<br>PRR: CGTTTCCCGCCTTCAGTTTAAA<br>PR: FAM-TCAGTGTTTGACACACACACTAAGCGTGCC-BHQ1	Mazzara M, Querci M, Bonfini L, Charels D, Cordeil S, Van den Eede G. Event-specific Method for the Quantification of Soybean Line MON 87701 Using Real-Time PCR. Protocol. 2011
Soybean<br><b>MON87708</b>	3'-border region between insert and soybean genome<br>Amplicon length: 91 b.p.	PRF: TCATACTCATTGCTGATCCATGTAG<br>PRR: AGAACAAATTAACGAAAAGACAGAACG<br>PR: FAM-TCCCGGACTTTAGCTCAAAATGCATGTA-BHQ1	Savini et al. Event-specific Method for the Quantification of Soybean MON87708 Using Real-time PCR. Validation Report and Validated Method. 2013
Soybean<br><b>MON89788</b>	5'-border region between insert and soybean genome<br>Amplicon length: 139 b.p.	PRF: TCCCGCTCTAGCGCTTCAAT<br>PRR: TCGAGCAGGACCTGCAGAA<br>PR: ROX-CTGAAGGCGGGAAACGACAATCTG-BHQ2	Mazzara M, Querci M, Bogni A, Grazioli E, Cordeil S, Van den Eede G. Event-specific Method for the Quantification of Soybean Line MON 89788 Using Real-Time PCR. Protocol. 2008
Soybean<br><b>SYHT0H2</b>	3'-border region between insert and soybean genome<br>Amplicon length: 88 b.p.	PRF: GGGAATTGGGTACCATGCC<br>PRR: TGTGTGCCATTGGTTTAGGGT<br>PR: ROX-CCAGCATGGCCGTATCCGCAA-BHQ2	Hart H. Event SYHT0H2 Soybean. Real-time, Event-specific Polymerase Chain Reaction Method. Assessment. 2012

Object

Target

Oligonucleotides

Refernce

Maize 176

3'-border region between insert and maize genome Amplicon length: 82 b.p.

PRF: GGCCGTGAACGAGCTGTT PRR: GGGAAGAAGCCTACATGTTTTCTAA PR: FAM-AGCAACCAGATCGGCCGACACC-BHQ1

Savini C, Bonfini L, Querci M, Mazzara M, Cordeil S, Van den Eede G. Event-specific Method for the Quantitation of Maize Line Bt176 Using Real-Time PCR - Validation Report and Protocol. 2011.

Maize 3272

5'-border region between insert and maize genome Amplicon length: 95 b.p.

PRF: TCATCAGACCAGATTCTCTTTTATGG PRR: CGTTTCCCGCCTTCAGTTTA PR: FAM-ACTGCTGACGCGGCCAAACACTG-BHQ1

Charles Delobel C, Foti N, Mazzara M, Van den Eede G. Event-specific Method for the Quantitation of Maize Event 3272 Using Real-Time PCR - Validation Report and Protocol. 2008. JRC48922

Maize 5307

3'-border region between insert and maize genome Amplicon length: 107 b.p.

PRF: CATGGCCGTATCCGCAATGTG PRR: TGCACCCTTTGCCAGTGG PR: FAM-ACCACAATATACCCTCTTCCCTGGGCCAG-BHQ1

Hart H. Event 5307 Maize. Real-time, Event-specific Polymerase Chain Reaction Method. Annex B. 2011

Maize Bt11

3'-border region between insert and maize genome Amplicon length: 70 b.p.

PRF: GCGGAACCCCTATTTGTTTA PRR: TCCAAGAATCCCTCCATGAG PR: ROX-AAATACATTCAAATATGTATCCGCTCA-BHQ2

Mazzara M, Puumalaainen J, Van den Eede G. Validation of the GMO Specific Detection Method Developed by NVI/NRA for Bt11 in Sweet Corn Maize - Validation Report and Protocol. 2005. JRC32111

Maize GA21

5'-border region between insert and maize genome Amplicon length: 112 b.p.

PRF: CTTATCGTTATGCTATTTGCAACTTTAGA PRR: TGGCTCGCGATCCTCCT PR: ROX-CATATACTAACTCATATCTCTTTCTCAACAGCAGGTGGGT-BHQ2

Paoletti C, Mazzara M, Puumalaainen J, Rasulo D, Van den Eede G. Event-specific Method for the Quantitation of Maize Line GA21 Using Real-Time PCR - Validation Report and Protocol. 2005. JRC32087

Maize LY038

5'-border region between insert and maize genome Amplicon length: 112 b.p.

PRF: TGGGTTCAGTCTGCGAATGTT PRR: AGGAATTCGATATCAAGCTTATCGA PR: FAM-CGAGCGGAGTTTATGGGTCGACGG-BHQ1

Charles Delobel C, Grazioli E, Larcher S, Mazzara M, Van den Eede G. Event-specific Method for the Quantification of Maize Line LY038 Using Real-time PCR - Validation Report and Protocol. EUR 23647 EN. Luxembourg (Luxembourg): Publications Office of the European Union, 2008. JRC48919

Maize MIR162

3'-border region between insert and maize genome Amplicon length: 92 b.p.

PRF: GCGCGGTGTCATCTATGTTACTAG PRR: TGCCTTATCTGTTGCCTTCAGA PR: FAM-TCTAGACAATTCAGTACATTAAAAACGTCCGCCA-BHQ1

Charles Delobel C, Mazzara M, Bevilacqua A, Van den Eede G. Event-specific Method for the Quantitation of Maize MIR162 by Real-time PCR. Validation Report and Protocol. 2011. JRC68155

Maize MIR604

5'-border region between insert and maize genome Amplicon length: 76 b.p.

PRF: GCGCACGCAATTCAACAG PRR: GGTCATAACGTGACTCCCTTAATTCT PR: ROX-AGGCGGGAAACGACAATCTGATCATG-BHQ2

Mazzara M, Munaro B, Foti N, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Maize Line MIR604 Using Real-Time PCR - Validation Report and Protocol. 2007. JRC37490

Maize MON810

5'-border region between insert and maize genome Amplicon length: 92 b.p.

PRF: TCGAAGGACGAAGGACTCTAACGT PRR: GCCACCTTCCTTTTCCACTATCTT PR: FAM-AACATCCTTTGCCATTGCCCAGC-BHQ1

ISO 21569:2005: Foodstuffs-Methods of analysis for the detection of genetically modified organisms and derived products-Qualitative nucleic acid based methods

Maize MON863

5'-border region between insert and maize genome Amplicon length: 84 b.p.

PRF: TGTTACGGCCTAAATGCTGAACT PRR: GTAGGATCGGAAAGCTTGGTAC PR: ROX-TGAACACCCATCCGAACAAGTAGGGTCA-BHQ2

Mazzara M, Foti N, Price S, Paoletti C, Van den Eede G. Event-specific Method for the Quantitation of Maize Line MON 863 Using Real-Time PCR - Validation Report and Protocol. 2005. JRC32105

Maize MON88017

3'-border region between insert and maize genome Amplicon length: 94 b.p.

PRF: GAGCAGGACCTGCAGAAGCT PRR: TCCGGAGTTGACCATCCA PR: ROX-TCCCGCCTTCAGTTTAAACAGAGTCGGGT-BHQ2

Charles Delobel C, Foti N, Grazioli E, Mazzara M, Van den Eede G. Event-specific Method for the Quantitation of Maize Line MON 88017 Using Real-Time PCR - Validation Report and Protocol. 2008. JRC48920

Maize MON89034

3'-border region between insert and maize genome Amplicon length: 77 b.p.

PRF: TTCTCCATATTGACCATCATACTCATT PRR: CGGTATCTATAATACCGTGGTTTTTAAA PR: FAM–ATCCCCGGAAATTATGTT–MGBNFQ

Savini C, Bogni A, Grazioli E, Munaro B, Mazzara M, Van den Eede G. Event-specific Method for the Quantitation of Maize Line MON 89034 Using Real-Time PCR - Validation Report and Protocol. 2008. JRC48921

Maize MZIR098

5'-border region between insert and maize genome Amplicon length: 73 b.p.

PRF: ATCTCAGACACCAAACCGAGATC PRR: ACACCGTTAGGCTAGTGCCAGT PR: FAM-CAAGTGACAGCGAACGGAGCTGGTTT-BHQ1

European Union Reference Laboratory for GM Food and Feed (EURL GMFF), Joint Research Centre (JRC), European Commission. Event-specific Method for the Quantification of Maize MZIR098 Real-time PCR - Validation Report. 2018. JRC 114043. Online Publication

Maize MZHG0JG

3'-border region between insert and maize genome Amplicon length: 153 b.p.

PRF: TTAGCTAACGGCCAGGATC PRR: CACCGTGACATGCTTAGCA PR: FAM-CGCGTGAGCCTTTAGCAACTAGC-BHQ1

GenBit

Maize NK603

3'-border region between insert and maize genome Amplicon length: 108 b.p.

PR: ATGAATGACCTCGAGTAAGCTTGTTAA PRR: AAGAGATAACAGGATCCACTCAAACACT PR: ROX-TGGTACCACGCGACACACTTCCACTC-BHQ2

Mazzara M, Paoletti C, Puumalaainen J, Rasulo D, Van den Eede G. Event-specific Method for the Quantitation of Maize Line NK603 Using Real-Time PCR - Validation Report and Protocol. 2005. JRC32103

Maize T25

3'-border region between insert and maize genome Amplicon length: 102 b.p.

PRF: ACAAGCGTGTCGTGCTCCAC PRR: GACATGATACTCCTTCCACCG PR: ROX-TCATTGAGTCGTTCCGCCATTGTCG-BHQ2

Mazzara M, Grazioli E, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Maize Line T25 Using Real-Time PCR v. 1.01 - Validation Report and Validated Method. 2013. JRC84152

Maize TC1507

3'-border region between insert and maize genome Amplicon length: 58 b.p.

PRF: TAGTCTTCGGCCAGAATGG PRR: CTTTGCCAAGATCAAGCG PR: FAM-TAACTCAAGGCCCTCACTCCG-BHQ1

Mazzara M. Foti N, Price S, Paoletti C, Van den Eede G. Event-specific Method for the Quantitation of Maize Line TC1507 Using Real-Time PCR - Validation Report and Protocol. 2005. JRC32120

Maize DAS-40278-9

5'-border region between insert and maize genome Amplicon length: 98 b.p.

PRF: CACGAACCATTGAGTTACAATC PRR: TGGTTCATTGTATTCTGGCTTTG PR: FAM-CGTAGCTAACCTTCATTGTATTCCG-BHQ1

Savini C, Bogni A, Foti N, Mazzara M, Kreysa J. Event-specific Method for the Quantification of Maize DAS-40278-9 by Real-time PCR - Validation Report and Validated Method - Sampling and DNA Extraction of Maize TC15O7. EUR 25585 EN. Luxembourg (Luxembourg): Publications Office of the European Union, 2012

Rapeseed GT73 (RT73)

3'-border region between insert and rapeseed genome Amplicon length: 108 b.p.

PRF: CCATATTGACCATCATACTCATTGCT PRR: GCTTATACGAAGGCAAGAAAAGGA PR: FAM-TTCCCGGACATGAAGATCATCCTCCTT-BHQ1

Mazzara M, Grazioli E, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Oilseed Rape Line RT73 Using Real-Time PCR - Validation Report and Protocol. 2007. JRC37550

Rice LLRICE62

3'-border region between insert and rice genome Amplicon length: 88 b.p.

PRF: AGCTGGCGTAATAGCGAAGAGG PRR: TGCTAACGGGTGCATCGTCTA PR: FAM-CGCACCGATTATTTATACTTTTAGTCCACCT-BHQ1

Mazzara M, Grazioli E, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Rice Line LLRICE62 Using Real-Time PCR - Validation Report and Protocol. 2006. JRC34091

Sugar beet H7-1

5'-border region between insert and sugar beet genome Amplicon length: 108 b.p.

PRF: TGGGATCTGGGTGGCTCTAACT PRR: AATGCTGCTAAATCCTGAG PR: FAM-AAGGCGGGAAACGACAATCT-BHQ1

Mazzara M, Foti N, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Sugar beet line H7-1 Using Real-Time PCR v. 1.01 - Validation Report and Valideted Method. 2013. JRC84121

Soybean A2704-12

region containing inverted 3'- and 5'-sequences of bla gene Amplicon length: 64 b.p.

PRF: GCAAAAAAGCGGTTAGCTCCT PRR: ATTCAGGCTGCGCAACTGTT PR: ROX-GGAAGGGCGATCGGAGGACCG-BHQ2

Mazzara M, Querci M, Delobel C, Moens W, Cordeil S, Van den Eede G. Event-specific Method for the Quantification of Soybean Line A2704-12 Using Real-Time PCR. Protocol. 2007

Soybean A5547-127

5'-border region between insert and soybean genome Amplicon length: 71 b.p.

PRF: CTATTTGGTGGCATTTTTCC PRR: TGCGGCCAACTTACTTC PR: FAM-ACAACGATGACGGTATGACATTGCGG-BHQ1

Mazzara M, Querci M, Luque-Perez E, Ermolli M, Cordeil S, Van den Eede G. Event-specific Method for the Quantification of Soybean Event A5547-127 Using Real-Time PCR. Protocol. 2009

Soybean BPS CV-127-9

3'-border region between insert and soybean genome Amplicon length: 88 b.p.

PRF: AACAGAAGTTTCCGTTGAGCTTTAAGAC PRR: CATTCGTAGCTCGGATCGTGTAC PR: FAM-TTTGGGGAAGCTGTCCCATGCCC-BHQ1

Mazzara M, Querci M, Savini C, Bonfini L, Cordeil S, Van den Eede G. Event-specific Method for the Quantification of Soybean CV127 Using Real-Time PCR. Protocol. 2011

Soybean DP305423-1

3'-border region between insert and soybean genome Amplicon length: 93 b.p.

PRF: CGTGTTCTCTTTTTGGCTAGC PRR: GTGACCAATGAATACATAACACAAACTA PR: FAM-TGACACAAATGATTTTCATACAAAAGTCGAGA-BHQ1

Mazzara M, Munaro B, Delobel C, Grazioli E, Savini C, Van den Eede G. Event-specific Method for the Quantification of Soybean Event DP-305423-1 Using Real-Time PCR v. 1.01. Validation Report and Validated Method. 2013

Soybean FG72

3'-border region between insert and soybean genome Amplicon length: 70 b.p.

PRF : AGATTTGATCGGGCTGCAGG PRR: GCACGTATTGATGACCGCATTA PR: FAM–AATGTGGTTCATCCGTCTT–MGBNFQ

Savini et al. Event-specific Method for the Quantification of Soybean FG72 Using Real-Time PCR. Validation Report and Protocol. 2012

Soybean GTS 40-3-2

5'-border region between insert and soybean genome Amplicon length: 84 b.p.

PRF: TTCATTCAAAATAAGATCATACATACAGGTT PRR: GGCATTTGTAGGAGCCACCTT PR: MGB FAM–CCTTTTCCATTTGGG–MGBNFQ

Mazzara M, Querci M, Moens W, Grazioli E, Cordeil S, Van den Eede G. Event-specific Method for the Quantification of Soybean Line 40-3-2 Using Real-Time PCR. Protocol. 2009.

Soybean MON87701

5'-border region between insert and soybean genome Amplicon length: 89 b.p.

PRF: TGGTGATATGAAGATACATGCTTAGCAT PRR: CGTTTCCCGCCTTCAGTTTAAA PR: FAM-TCAGTGTTTGACACACACACTAAGCGTGCC-BHQ1

Mazzara M, Querci M, Bonfini L, Charels D, Cordeil S, Van den Eede G. Event-specific Method for the Quantification of Soybean Line MON 87701 Using Real-Time PCR. Protocol. 2011

Soybean MON87708

3'-border region between insert and soybean genome Amplicon length: 91 b.p.

PRF: TCATACTCATTGCTGATCCATGTAG PRR: AGAACAAATTAACGAAAAGACAGAACG PR: FAM-TCCCGGACTTTAGCTCAAAATGCATGTA-BHQ1

Savini et al. Event-specific Method for the Quantification of Soybean MON87708 Using Real-time PCR. Validation Report and Validated Method. 2013

Soybean MON89788

5'-border region between insert and soybean genome Amplicon length: 139 b.p.

PRF: TCCCGCTCTAGCGCTTCAAT PRR: TCGAGCAGGACCTGCAGAA PR: ROX-CTGAAGGCGGGAAACGACAATCTG-BHQ2

Mazzara M, Querci M, Bogni A, Grazioli E, Cordeil S, Van den Eede G. Event-specific Method for the Quantification of Soybean Line MON 89788 Using Real-Time PCR. Protocol. 2008

Soybean SYHT0H2

3'-border region between insert and soybean genome Amplicon length: 88 b.p.

PRF: GGGAATTGGGTACCATGCC PRR: TGTGTGCCATTGGTTTAGGGT PR: ROX-CCAGCATGGCCGTATCCGCAA-BHQ2

Hart H. Event SYHT0H2 Soybean. Real-time, Event-specific Polymerase Chain Reaction Method. Assessment. 2012

Sense genes

Object	Target	Oligonucleotides	Refernce
Sense gene<br><b>bar</b>	region of phosphinothricin N-acetyltransferase gene from <i>Streptomyces hydroscopicus</i><br>Amplicon length: 60 b.p.	PRF: ACAAGCACGGTCAACTTCC<br>PRR: GAGGTCGTCCGTCCACTC<br>PR: FAM-TACCGAGCCGCAGGAACC-BHQ1	Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011
Sense gene<br><a href=#popBox_stacks_in_4921_page15 class=call-modal><b>cp4 epsps1</b></a>	region of 5-enolpyruvylshikimate-3-phosphate synthase gene from <i>Agrobacterium tumefaciens</i><br>Amplicon length: 226 b.p.	PRF: GCATGCTTCACGGTGCAA<br>PRR1: TGAAGGACCTGTGGGAGAT<br>PRR2: TGAAGGACCGGTGGGAGAT<br>PR: FAM-CGCAACCGCCCGCAAATC-BHQ1	E. Barbau-Piednoir, P. Stragier, N. Roosens, M. Mazzara, C. Savini, G. Van den Eede, M. Van den Bulcke. Inter-laboratory Testing of GMO Detection by Combinatory SYBR Green PCR Screening (CoSYPS). Food Anal. Methods. 2014 / GenBit
Sense gene<br><a href=#popBox_stacks_in_4921_page15 class=call-modal><b>cp4 epsps2</b></a>	region of 5-enolpyruvylshikimate-3-phosphate synthase gene from <i>Agrobacterium tumefaciens</i><br>Amplicon length: 88 b.p.	PRF: GGGATGACGTTAATTGGCTCTG<br>PRR: GGCTGCTTGCACCGTGAAG<br>PR: FAM-CACGCCGTGGAAACAGAAGACATGACC-BHQ1	GenBit
Sense gene<br><b>mepsps</b>	region of modified 5-enolpyruvylshikimate-3-phosphate synthase gene from <i>Zea mays</i><br>Amplicon length: 112 b.p.	PRF: GTCGAAGCGGACAAAGCTG<br>PRR: CTGGGGGATCCACTAGTTCT<br>PR: ROX-CTGTAGTTGTTGGCTGTGGTGGAA-BHQ2	GenBit
Sense gene<br><b>cry1A.105</b>	region of Cry1A.105 delta endotoxin gene from <i>Bacillus thuringiensis</i><br>Amplicon length: 133 b.p.	PRF: TCAGAGGTCCAGGGTTTACAGG<br>PRR: GTAGTAGAGGCATAGCGGATTCTTG<br>PR: FAM-TCCACTTGTGCGACGAAGAATGTCT-BHQ1	Dinon A. et al. Development and validation of real-time PCR screening methods for detection of cry1A.105 and cry2Ab2 genes in genetically modified organisms // Anal Bioanal Chem (2011), 400:1433-1442
Sense gene<br><b>cry1Ab</b>	region of Cry1Ab1 delta endotoxin gene from <i>Bacillus thuringiensis</i><br>Amplicon length: 77 b.p.	PRF: CAGTTCCTGCTSAGCGAGTT<br>PRR: CCRAAGATGCCCCAGATG<br>PR: FAM-ATGTCCACSAGGCCCAGSACG-BHQ1	GenBit
Sense gene<br><b>cry1Ac</b>	region of Cry1Ac delta endotoxin gene from <i>Bacillus thuringiensis</i><br>Amplicon length: 137 b.p.	PRF: TACTTGGTGGAGAACGCATT<br>PRR: AAAGATACCCCAGATGATGTCA<br>PR: FAM-CACCTGGCACGAACTCGCTGAG-BHQ1	Dong et al. GMDD: a database of GMO detection methods // BMC Bioinformatics (2008), 9:260
Sense gene<br><b>cry1F</b>	region of Cry1F delta endotoxin gene from <i>Bacillus thuringiensis</i><br>Amplicon length: 245 b.p.	PRF: CATCTGATTGGAGCCTCTTTCTT<br>PRR: GGTAAGGGTGAAGTTGTTGATGG<br>PR: FAM-CGCACATCTTCTCTCAGTTGGGCA-BHQ1	GenBit
Sense gene<br><b>cry2Ab</b>	region of Cry2Ab delta endotoxin gene from <i>Bacillus thuringiensis</i><br>Amplicon length: 120 b.p.	PRF: TTCTAACTACTTCCCCGACTACTTC<br>PRR: GACGGAGAGGCGATGTTC<br>PR: ROX-TCTCTGGTGTTCCTCTCGTCGTCCGCA-BHQ2	Dinon A. et al. Development and validation of real-time PCR screening methods for detection of cry1A.105 and cry2Ab2 genes in genetically modified organisms // Anal Bioanal Chem (2011), 400:1433-1442
Sense gene<br><b>cry3A</b>	region of Cry3A delta endotoxin gene from <i>Bacillus thuringiensis</i><br>Amplicon length: 147 b.p.	PRF: CGACAGTTCTACCACTAAGGAT<br>PRR: TTTGTATAGAAGCTCACGAGGG<br>PR: FAM-CCAGAAGGGTATCTCCGTTGTG-BHQ1	GenBit
Sense gene<br><b>mcry3A</b>	region of modified cry3A delta endotoxin gene from <i>Bacillus thuringiensis</i><br>Amplicon length: 154 b.p.	PRF: AACAACACCGAGGCCCTG<br>PRR: CAGCAGCACCACCAAGGACG<br>PR: FAM-CACCACGCTGATGCCCT-BHQ1	GenBit
Sense gene<br><b>cry34Ab1</b>	region of cry34Ab1 delta endotoxin gene from <i>Bacillus thuringiensis</i><br>Amplicon length: 167 b.p.	PRF: CAAGACCAAGCTCGACGG<br>PRR: GGTTGTCGAAGTAGAGGCTGAT<br>PR: ROX-CGCACCTCCCCGACCAAC-BHQ2	GenBit
Sense gene<br><b>cry35Ab1</b>	region of cry34Ab1 delta endotoxin gene from <i>Bacillus thuringiensis</i><br>Amplicon length: 78 b.p.	PRF: CCATGCTCGACACCAACAA<br>PRR: CGAGGGAGAGGTAGGTGGC<br>PR: ROX-TGTACGAGATCAGCAACCACGCC-BHQ2	GenBit
Sense gene<br><b>cry3Bb1</b>	region of cry3Bb1 delta endotoxin gene from <i>Bacillus thuringiensis</i><br>Amplicon length: 89 b.p.	PRF: GAGCACGACACGATCAAGGT<br>PRR: AGGGTGGAGTTGGGGTTGT<br>PR: FAM-CAGCGGGTACTGGTTGTGGTTG-BHQ1	Jia et al. Screening genetically modified organisms using multiplex-PCR coupled with oligonucleotide microarray // Biosens. Bioelctron. 2006, 22(1): 71-77
Sense gene<br><b>cry9C</b>	region of cry9C delta endotoxin gene from <i>Bacillus thuringiensis</i><br>Amplicon length: 198 b.p.	PRF: CTGCTGCTGCTGAAGGATGC<br>PRR: CCTGCGGAACTGGTGGTAG<br>PR: ROX-CGGCGAGGGCTGGGGCTT-BHQ2	Jia et al. Screening genetically modified organisms using multiplex-PCR coupled with oligonucleotide microarray // Biosens. Bioelctron. 2006, 22(1): 71-77
Sense gene<br><b>gox/gox v247</b>	region of glyphosate oxidase gene from <i>Ochrobactrum anthropi</i></i><br>Amplicon length: 99 b.p.	PRF: ATGCTAGCCACCTTATCCG<br>PRR: TGAGACGACGAAGTTCCCA<br>PR: ROX-TCACCTTACCGTGTACCGTGGA-BHQ2	GenBit
Sense gene<br><b>pat</b>	region of phosphinothricin N-acetyltransferase gene from <i>Streptomyces viridochromogenes</i><br>Amplicon length: 68 b.p.	PRF: TTGAGGGTGTTGTGGCTGGTA<br>PRR: TGTCCAATCGTAAGCGTTCCT<br>PR: ROX-CTTCCAGGGCCCAGCGTAAGCA-BHQ2	Weighardt F, Barbati C, Paoletti C, Querci M, Kay S, De Beuckeleer M, Van den Eede G. Real-Time Polymerase Chain Reaction-Based Arrpoach for Quantification of the pat Gene in the T25 Zea mays Event' Journal OF AOAC International, Vol. 87, No. 6, p. 1342-1355
Sense gene<br><b>CP-PRSV-P</b>	region of CI gene, coat protein of papaya ring spot virus, strain P<br>Amplicon length: 133 b.p.	PRF: CGACTTTGACATCACTGGTGA<br>PRR: CTCAACGAATGTTCCTCCAAT<br>PR: FAM-TGGACCTATAGATGTCACGTTCGAGA-BHQ1	GenBit

Object

Target

Oligonucleotides

Refernce

Sense gene bar

region of phosphinothricin N-acetyltransferase gene from Streptomyces hydroscopicus Amplicon length: 60 b.p.

PRF: ACAAGCACGGTCAACTTCC PRR: GAGGTCGTCCGTCCACTC PR: FAM-TACCGAGCCGCAGGAACC-BHQ1

Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011

Sense gene <a href=#popBox_stacks_in_4921_page15 class=call-modal>cp4 epsps1</a>

region of 5-enolpyruvylshikimate-3-phosphate synthase gene from Agrobacterium tumefaciens Amplicon length: 226 b.p.

PRF: GCATGCTTCACGGTGCAA PRR1: TGAAGGACCTGTGGGAGAT PRR2: TGAAGGACCGGTGGGAGAT PR: FAM-CGCAACCGCCCGCAAATC-BHQ1

E. Barbau-Piednoir, P. Stragier, N. Roosens, M. Mazzara, C. Savini, G. Van den Eede, M. Van den Bulcke. Inter-laboratory Testing of GMO Detection by Combinatory SYBR Green PCR Screening (CoSYPS). Food Anal. Methods. 2014 / GenBit

Sense gene <a href=#popBox_stacks_in_4921_page15 class=call-modal>cp4 epsps2</a>

region of 5-enolpyruvylshikimate-3-phosphate synthase gene from Agrobacterium tumefaciens Amplicon length: 88 b.p.

PRF: GGGATGACGTTAATTGGCTCTG PRR: GGCTGCTTGCACCGTGAAG PR: FAM-CACGCCGTGGAAACAGAAGACATGACC-BHQ1

GenBit

Sense gene mepsps

region of modified 5-enolpyruvylshikimate-3-phosphate synthase gene from Zea mays Amplicon length: 112 b.p.

PRF: GTCGAAGCGGACAAAGCTG PRR: CTGGGGGATCCACTAGTTCT PR: ROX-CTGTAGTTGTTGGCTGTGGTGGAA-BHQ2

GenBit

Sense gene cry1A.105

region of Cry1A.105 delta endotoxin gene from Bacillus thuringiensis Amplicon length: 133 b.p.

PRF: TCAGAGGTCCAGGGTTTACAGG PRR: GTAGTAGAGGCATAGCGGATTCTTG PR: FAM-TCCACTTGTGCGACGAAGAATGTCT-BHQ1

Dinon A. et al. Development and validation of real-time PCR screening methods for detection of cry1A.105 and cry2Ab2 genes in genetically modified organisms // Anal Bioanal Chem (2011), 400:1433-1442

Sense gene cry1Ab

region of Cry1Ab1 delta endotoxin gene from Bacillus thuringiensis Amplicon length: 77 b.p.

PRF: CAGTTCCTGCTSAGCGAGTT PRR: CCRAAGATGCCCCAGATG PR: FAM-ATGTCCACSAGGCCCAGSACG-BHQ1

GenBit

Sense gene cry1Ac

region of Cry1Ac delta endotoxin gene from Bacillus thuringiensis Amplicon length: 137 b.p.

PRF: TACTTGGTGGAGAACGCATT PRR: AAAGATACCCCAGATGATGTCA PR: FAM-CACCTGGCACGAACTCGCTGAG-BHQ1

Dong et al. GMDD: a database of GMO detection methods // BMC Bioinformatics (2008), 9:260

Sense gene cry1F

region of Cry1F delta endotoxin gene from Bacillus thuringiensis Amplicon length: 245 b.p.

PRF: CATCTGATTGGAGCCTCTTTCTT PRR: GGTAAGGGTGAAGTTGTTGATGG PR: FAM-CGCACATCTTCTCTCAGTTGGGCA-BHQ1

GenBit

Sense gene cry2Ab

region of Cry2Ab delta endotoxin gene from Bacillus thuringiensis Amplicon length: 120 b.p.

PRF: TTCTAACTACTTCCCCGACTACTTC PRR: GACGGAGAGGCGATGTTC PR: ROX-TCTCTGGTGTTCCTCTCGTCGTCCGCA-BHQ2

Dinon A. et al. Development and validation of real-time PCR screening methods for detection of cry1A.105 and cry2Ab2 genes in genetically modified organisms // Anal Bioanal Chem (2011), 400:1433-1442

Sense gene cry3A

region of Cry3A delta endotoxin gene from Bacillus thuringiensis Amplicon length: 147 b.p.

PRF: CGACAGTTCTACCACTAAGGAT PRR: TTTGTATAGAAGCTCACGAGGG PR: FAM-CCAGAAGGGTATCTCCGTTGTG-BHQ1

GenBit

Sense gene mcry3A

region of modified cry3A delta endotoxin gene from Bacillus thuringiensis Amplicon length: 154 b.p.

PRF: AACAACACCGAGGCCCTG PRR: CAGCAGCACCACCAAGGACG PR: FAM-CACCACGCTGATGCCCT-BHQ1

GenBit

Sense gene cry34Ab1

region of cry34Ab1 delta endotoxin gene from Bacillus thuringiensis Amplicon length: 167 b.p.

PRF: CAAGACCAAGCTCGACGG PRR: GGTTGTCGAAGTAGAGGCTGAT PR: ROX-CGCACCTCCCCGACCAAC-BHQ2

GenBit

Sense gene cry35Ab1

region of cry34Ab1 delta endotoxin gene from Bacillus thuringiensis Amplicon length: 78 b.p.

PRF: CCATGCTCGACACCAACAA PRR: CGAGGGAGAGGTAGGTGGC PR: ROX-TGTACGAGATCAGCAACCACGCC-BHQ2

GenBit

Sense gene cry3Bb1

region of cry3Bb1 delta endotoxin gene from Bacillus thuringiensis Amplicon length: 89 b.p.

PRF: GAGCACGACACGATCAAGGT PRR: AGGGTGGAGTTGGGGTTGT PR: FAM-CAGCGGGTACTGGTTGTGGTTG-BHQ1

Jia et al. Screening genetically modified organisms using multiplex-PCR coupled with oligonucleotide microarray // Biosens. Bioelctron. 2006, 22(1): 71-77

Sense gene cry9C

region of cry9C delta endotoxin gene from Bacillus thuringiensis Amplicon length: 198 b.p.

PRF: CTGCTGCTGCTGAAGGATGC PRR: CCTGCGGAACTGGTGGTAG PR: ROX-CGGCGAGGGCTGGGGCTT-BHQ2

Jia et al. Screening genetically modified organisms using multiplex-PCR coupled with oligonucleotide microarray // Biosens. Bioelctron. 2006, 22(1): 71-77

Sense gene gox/gox v247

region of glyphosate oxidase gene from Ochrobactrum anthropi Amplicon length: 99 b.p.

PRF: ATGCTAGCCACCTTATCCG PRR: TGAGACGACGAAGTTCCCA PR: ROX-TCACCTTACCGTGTACCGTGGA-BHQ2

GenBit

Sense gene pat

region of phosphinothricin N-acetyltransferase gene from Streptomyces viridochromogenes Amplicon length: 68 b.p.

PRF: TTGAGGGTGTTGTGGCTGGTA PRR: TGTCCAATCGTAAGCGTTCCT PR: ROX-CTTCCAGGGCCCAGCGTAAGCA-BHQ2

Weighardt F, Barbati C, Paoletti C, Querci M, Kay S, De Beuckeleer M, Van den Eede G. Real-Time Polymerase Chain Reaction-Based Arrpoach for Quantification of the pat Gene in the T25 Zea mays Event' Journal OF AOAC International, Vol. 87, No. 6, p. 1342-1355

Sense gene CP-PRSV-P

region of CI gene, coat protein of papaya ring spot virus, strain P Amplicon length: 133 b.p.

PRF: CGACTTTGACATCACTGGTGA PRR: CTCAACGAATGTTCCTCCAAT PR: FAM-TGGACCTATAGATGTCACGTTCGAGA-BHQ1

GenBit

Regulatory and marker sequences

Object	Target	Oligonucleotides	Refernce
Promoter<br><b>p35S</b>	region of 35S promoter from cauliflower mosaic virus<br>Amplicon length: 102 b.p.	PRF: TCGTTGAAGATGCCTCTGC<br>PRR: CTTGCTTTGAAGACGTGGTT<br>PR: FAM-CAAAGATGGACCCCCACCCAC-BHQ1	Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011
Promoter<br><b>pFMV</b>	region of 35S promoter from figwort mosaic virus<br>Amplicon length: 78 b.p.	PRF: CAAAATAACGTGGAAAAGAGCT<br>PRR: TCTTTTGTGGTCGTCACTGC<br>PR: ROX-CTGACAGCCCACTCACTAATGC-BHQ2	Official Compendium of Analytical Methods according to § 64 LFGB (Food and Feed Law) (2014). Food Analysis, Detection of DNA sequence of the FMV-promoter (pFMV) in foodstuffs with real-time PCR. BLV L 00.00-148, Beuth, Berlin Koln
Promoter<br><b>pNOS</b>	region of nopaline synthase promoter from <i>Agrobacterium tumefaciens</i><br>Amplicon length: 79 b.p.	PRF: GTGACCTTAGGCGACTTTTGAAC<br>PRR: CGCGGGTTTCTGGAGTTTAA<br>PR: FAM-CGCAATAATGGTTTCTGACGTATGTGCTTAGC-BHQ1	Debode F., Janssen E., Berben G. Development of 10 new screening PCR assays for GMO detection targeting promoters (pFMV, pNOS, pSSuAra, pTA29, pUbi, pRice actin) and terminators (t35S, tE9, tOCS, tg7) // Eur Food Res Technol (2013) 236:659-669
Promoter<br><b>pRice_actin</b>	region of actin 1 promoter from <i>Oryza sativa</i><br>Amplicon length: 95 b.p.	PRF: TCGAGGTCATTCATATGCTTGAG<br>PRR: TTTTAACTGATGTTTTCACTTTTGACC<br>PR: FAM-AGAGAGTCGGGATAGTCCAAAATAAAACAAAGGTA-BHQ1	Debode F., Janssen E., Berben G. Development of 10 new screening PCR assays for GMO detection targeting promoters (pFMV, pNOS, pSSuAra, pTA29, pUbi, pRice actin) and terminators (t35S, tE9, tOCS, tg7) // Eur Food Res Technol (2013) 236:659-669
Promoter<br><b>pSSuAra</b>	region of Ribulose-1,5-bisphosphate carboxylase (RuBisCO) small subunit promoter from <i>Arabidopsis thaliana</i><br>Amplicon length: 95 b.p.	PRF: GGCCTAAGGAGAGGTGTTGAGA<br>PRR: CTCATAGATAACGATAAGATTCATGGAATT<br>PR: FAM-CCTTATCGGCTTGAACCGCTGGAATAA-BHQ1	Debode F., Janssen E., Berben G. Development of 10 new screening PCR assays for GMO detection targeting promoters (pFMV, pNOS, pSSuAra, pTA29, pUbi, pRice actin) and terminators (t35S, tE9, tOCS, tg7) // Eur Food Res Technol (2013) 236:659-669
Promoter<br><b>pTA29</b>	region of TA29 promoter from <i>Nicotiana tabacum</i><br>Amplicon length: 117 b.p.	PRF: GAAGCTGTGCTAGAGAAGATGTTTATTC<br>PRR: GCTCGAAGTATGCACATTTAGCAA<br>PR: FAM-AGTCCAGCCACCCACCTTATGCAAGTC-BHQ1	Debode F., Janssen E., Berben G. Development of 10 new screening PCR assays for GMO detection targeting promoters (pFMV, pNOS, pSSuAra, pTA29, pUbi, pRice actin) and terminators (t35S, tE9, tOCS, tg7) // Eur Food Res Technol (2013) 236:659-669
Promoter<br><b>pUbi</b>	region of ubiquitin promoter from <i>Zea mays</i><br>Amplicon length: 76 b.p.	PRF: GAGTAGATAATGCCAGCCTGTTAAAC<br>PRR: ACGCGACGCTGCTGGTT<br>PR: FAM-CGTCGACGAGTCTAACGGACACCAAC-BHQ1	Debode F., Janssen E., Berben G. Development of 10 new screening PCR assays for GMO detection targeting promoters (pFMV, pNOS, pSSuAra, pTA29, pUbi, pRice actin) and terminators (t35S, tE9, tOCS, tg7) // Eur Food Res Technol (2013) 236:659-669
Terminator<br><b>t35S</b>	region of 35S terminator from cauliflower mosaic virus<br>Amplicon length: 118 b.p.	PRF: AGGGTTTCTTATATGCTCAACACATG<br>PRR: TCACCAGTCTCTCTCTACAAATCTATCAC<br>PR: ROX-AAACCCTATAAGAACCCTAATTCCCTTATCTGGGA-BHQ2	Debode F., Janssen E., Berben G. Development of 10 new screening PCR assays for GMO detection targeting promoters (pFMV, pNOS, pSSuAra, pTA29, pUbi, pRice actin) and terminators (t35S, tE9, tOCS, tg7) // Eur Food Res Technol (2013) 236:659-669
Terminator<br><b>tE9</b>	region of Ribulose-1,5-bisphosphate carboxylase (RuBisCO) small subunit terminator from <i>Pisum sativum</i><br>Amplicon length: 87 b.p.	PRF: TGAGAATGAACAAAAGGACCATATCA<br>PRR: TTTTTATTCGGTTTTCGCTATCG<br>PR: ROX-TCATTAACTCTTCTCCATCCATTTCCATTTCACAGT-BHQ2	Debode F., Janssen E., Berben G. Development of 10 new screening PCR assays for GMO detection targeting promoters (pFMV, pNOS, pSSuAra, pTA29, pUbi, pRice actin) and terminators (t35S, tE9, tOCS, tg7) // Eur Food Res Technol (2013) 236:659-669
Terminator<br><b>tg7</b>	region of g7 terminator from <i>Agrobacterium tumefaciens</i><br>Amplicon length: 97 b.p.	PRF: ATGCAAGTTTAAATTCAGAAATATTTCAA<br>PRR: ATGTATTACACATAATATCGCACTCAGTCT<br>PR: ROX-ACTGATTATATCAGCTGGTACATTGCCGTAGATGA-BHQ2	Debode F., Janssen E., Berben G. Development of 10 new screening PCR assays for GMO detection targeting promoters (pFMV, pNOS, pSSuAra, pTA29, pUbi, pRice actin) and terminators (t35S, tE9, tOCS, tg7) // Eur Food Res Technol (2013) 236:659-669
Terminator<br><b>tNOS</b>	region of nopaline synthase terminator from <i>Agrobacterium tumefaciens</i><br>Amplicon length: 98 b.p.	PRF: ATGCTTAACGTAATTCAACAGA<br>PRR: ATCGTTCAAACATTTGGCAAT<br>PR: FAM-ATCGCAAGACCGGCAACAGGATTC-BHQ1	Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011
Terminator<br><b>tOCS</b>	region of octopine synthase terminator from <i>Agrobacterium tumefaciens</i><br>Amplicon length: 85 b.p.	PRF: CGGTCAAACCTAAAAGACTGATTACA<br>PRR: CGCTCGGTGTCGTAGATACT<br>PR: ROX-TCTTATTCAAATTTCAAAAGTGCCCCAGGG-BHQ2	Debode F., Janssen E., Berben G. Development of 10 new screening PCR assays for GMO detection targeting promoters (pFMV, pNOS, pSSuAra, pTA29, pUbi, pRice actin) and terminators (t35S, tE9, tOCS, tg7) // Eur Food Res Technol (2013) 236:659-669
Terminator<br><b>tORF25</b>	region of ORF25 terminator from <i>Agrobacterium tumefaciens</i><br>Amplicon length: 165 b.p.	PRF: ATCAGATTATTGACTGTCATTTGT <br>PRR: ATAATGTTGTCGGTATTTTGTAA <br>PR: FAM-CATATAGATTTTCACTGTGCGACGC-BHQ1	GenBit
Selective marker<br><b>bla</b>	region of beta-lactamase <i>Escherichia coli</i><br>Amplicon length: 85 b.p.	PRF: CGTTTGGTATGGCTTCATTCA<br>PRR: CTAACCGCTTTTTTGCACAAC<br>PR: FAM-CCGGTTCCCAACGATCAAGG-BHQ1	Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011
Selective marker<br><b>GUS</b>	region of beta-glucuronidase gene from <i>Escherichia coli</i><br>Amplicon length: 82 b.p.	PRF: TTTCTTTAACTATGCCGGAATCCATC<br>PRR: CACCACGGTGATATCGTCCAC<br>PR: FAM-AATGCTCTACACCACGCCGAACACCTG-BHQ1	GenBit
Selective marker<br><b>nptII</b>	region of neomycin phosphotransferase II gene from <i>Escherichia coli</i><br>Amplicon length: 210 b.p.	PRF: TCACCTTGCTCCTGCC<br>PRR: CCCCTGATGCTCTTCG<br>PR: ROX-ATGCAATGCGGCGGCTG-BHQ2	Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011
Selective marker<br><b>pmi</b>	region of phosphomannose isomerase gene from <i>Escherichia coli</i> <br>Amplicon length: 149 b.p.	PRF: GCGAACTGCCTTTCCTGT <br>PRR: ATCTTTATAGTTACGCTCGGC <br>PR: FAM-TCTCCATTCAGGTTCATCCAAACAAACA-BHQ1	GenBit

Object

Target

Oligonucleotides

Refernce

Promoter p35S

region of 35S promoter from cauliflower mosaic virus Amplicon length: 102 b.p.

PRF: TCGTTGAAGATGCCTCTGC PRR: CTTGCTTTGAAGACGTGGTT PR: FAM-CAAAGATGGACCCCCACCCAC-BHQ1

Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011

Promoter pFMV

region of 35S promoter from figwort mosaic virus Amplicon length: 78 b.p.

PRF: CAAAATAACGTGGAAAAGAGCT PRR: TCTTTTGTGGTCGTCACTGC PR: ROX-CTGACAGCCCACTCACTAATGC-BHQ2

Official Compendium of Analytical Methods according to § 64 LFGB (Food and Feed Law) (2014). Food Analysis, Detection of DNA sequence of the FMV-promoter (pFMV) in foodstuffs with real-time PCR. BLV L 00.00-148, Beuth, Berlin Koln

Promoter pNOS

region of nopaline synthase promoter from Agrobacterium tumefaciens Amplicon length: 79 b.p.

PRF: GTGACCTTAGGCGACTTTTGAAC PRR: CGCGGGTTTCTGGAGTTTAA PR: FAM-CGCAATAATGGTTTCTGACGTATGTGCTTAGC-BHQ1

Debode F., Janssen E., Berben G. Development of 10 new screening PCR assays for GMO detection targeting promoters (pFMV, pNOS, pSSuAra, pTA29, pUbi, pRice actin) and terminators (t35S, tE9, tOCS, tg7) // Eur Food Res Technol (2013) 236:659-669

Promoter pRice_actin

region of actin 1 promoter from Oryza sativa Amplicon length: 95 b.p.

PRF: TCGAGGTCATTCATATGCTTGAG PRR: TTTTAACTGATGTTTTCACTTTTGACC PR: FAM-AGAGAGTCGGGATAGTCCAAAATAAAACAAAGGTA-BHQ1

Promoter pSSuAra

region of Ribulose-1,5-bisphosphate carboxylase (RuBisCO) small subunit promoter from Arabidopsis thaliana Amplicon length: 95 b.p.

PRF: GGCCTAAGGAGAGGTGTTGAGA PRR: CTCATAGATAACGATAAGATTCATGGAATT PR: FAM-CCTTATCGGCTTGAACCGCTGGAATAA-BHQ1

Promoter pTA29

region of TA29 promoter from Nicotiana tabacum Amplicon length: 117 b.p.

PRF: GAAGCTGTGCTAGAGAAGATGTTTATTC PRR: GCTCGAAGTATGCACATTTAGCAA PR: FAM-AGTCCAGCCACCCACCTTATGCAAGTC-BHQ1

Promoter pUbi

region of ubiquitin promoter from Zea mays Amplicon length: 76 b.p.

PRF: GAGTAGATAATGCCAGCCTGTTAAAC PRR: ACGCGACGCTGCTGGTT PR: FAM-CGTCGACGAGTCTAACGGACACCAAC-BHQ1

Terminator t35S

region of 35S terminator from cauliflower mosaic virus Amplicon length: 118 b.p.

PRF: AGGGTTTCTTATATGCTCAACACATG PRR: TCACCAGTCTCTCTCTACAAATCTATCAC PR: ROX-AAACCCTATAAGAACCCTAATTCCCTTATCTGGGA-BHQ2

Terminator tE9

region of Ribulose-1,5-bisphosphate carboxylase (RuBisCO) small subunit terminator from Pisum sativum Amplicon length: 87 b.p.

PRF: TGAGAATGAACAAAAGGACCATATCA PRR: TTTTTATTCGGTTTTCGCTATCG PR: ROX-TCATTAACTCTTCTCCATCCATTTCCATTTCACAGT-BHQ2

Terminator tg7

region of g7 terminator from Agrobacterium tumefaciens Amplicon length: 97 b.p.

PRF: ATGCAAGTTTAAATTCAGAAATATTTCAA PRR: ATGTATTACACATAATATCGCACTCAGTCT PR: ROX-ACTGATTATATCAGCTGGTACATTGCCGTAGATGA-BHQ2

Terminator tNOS

region of nopaline synthase terminator from Agrobacterium tumefaciens Amplicon length: 98 b.p.

PRF: ATGCTTAACGTAATTCAACAGA PRR: ATCGTTCAAACATTTGGCAAT PR: FAM-ATCGCAAGACCGGCAACAGGATTC-BHQ1

Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011

Terminator tOCS

region of octopine synthase terminator from Agrobacterium tumefaciens Amplicon length: 85 b.p.

PRF: CGGTCAAACCTAAAAGACTGATTACA PRR: CGCTCGGTGTCGTAGATACT PR: ROX-TCTTATTCAAATTTCAAAAGTGCCCCAGGG-BHQ2

Terminator tORF25

region of ORF25 terminator from Agrobacterium tumefaciens Amplicon length: 165 b.p.

PRF: ATCAGATTATTGACTGTCATTTGT PRR: ATAATGTTGTCGGTATTTTGTAA PR: FAM-CATATAGATTTTCACTGTGCGACGC-BHQ1

GenBit

Selective marker bla

region of beta-lactamase Escherichia coli Amplicon length: 85 b.p.

PRF: CGTTTGGTATGGCTTCATTCA PRR: CTAACCGCTTTTTTGCACAAC PR: FAM-CCGGTTCCCAACGATCAAGG-BHQ1

Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011

Selective marker GUS

region of beta-glucuronidase gene from Escherichia coli Amplicon length: 82 b.p.

PRF: TTTCTTTAACTATGCCGGAATCCATC PRR: CACCACGGTGATATCGTCCAC PR: FAM-AATGCTCTACACCACGCCGAACACCTG-BHQ1

GenBit

Selective marker nptII

region of neomycin phosphotransferase II gene from Escherichia coli Amplicon length: 210 b.p.

PRF: TCACCTTGCTCCTGCC PRR: CCCCTGATGCTCTTCG PR: ROX-ATGCAATGCGGCGGCTG-BHQ2

Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011

Selective marker pmi

region of phosphomannose isomerase gene from Escherichia coli Amplicon length: 149 b.p.

PRF: GCGAACTGCCTTTCCTGT PRR: ATCTTTATAGTTACGCTCGGC PR: FAM-TCTCCATTCAGGTTCATCCAAACAAACA-BHQ1

GenBit

Species identity

Object	Target	Oligonucleotides	Refernce
Species belonging:<br><b>Potato</b><br><i>(Solanum tuberosum)</i>	region of UDP-glucose pyrophosphorylase gene (UGPase)<br>Amplicon length: 89 b.p.	PRF: GGACATGTGAAGAGACGGAGC<br>PRR: CCTACCTCTACCCCTCCTA<br>PR: ROX-TGAGGAGGTGCGAGGTAATGGTGGTAG-BHQ2	Savini C, Foti N, Mazzara M, Charles Delobel C, Van den Eede G. Event-specific Method for the Quantitation of Event EH92-527-1 Potato Using Real-Time PCR - Validation Report and Protocol. 2006. JRC34758
Species belonging:<br><b>Maize</b><br><i>(Zea mays)</i>	region of zein gene</i><br>Amplicon length: 82 b.p.	PRF: TTACCGCTTCAGACGATGCC<br>PRR: CATTATTTGCGACACATGCTC<br>PR: FAM-TGACGCCTAACATGATGTCACCA-BHQ1	Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011
Species belonging:<br><b>Flax</b><br><i>(Linum </i>L.<i>)</i>	SAD gene region<br>Amplicon length: 68 b.p.	PRF: GCTCCACCCAGTCACCACCT<br>PRR: TGCGAGGAGATCTGGAGGAG<br>PR: ROX-TGTTGAGGGAGCGTGTTGAAGGGA-BHQ2	Rosario V. et al. Development of an event-specific assay for the qualitative and quantitative detection of the genetically modified flax CDC Triffid (FP967) // Food Control, 2014, 41:128-133
Species belonging:<br><b>Sunflower</b><br><i>(Helianthus annuus)</i>	TUB1 gene region<br>Amplicon length: 128 b.p.	PRF: CGTAGCATATCGTTTCACCG<br>PRR: TCAAGGCAATAGAGATTCCCAA<br>PR: ROX-AGGAAAAATGAGAGAAATCGTAAGCA-BHQ2	GenBit
Species belonging:<br><b>Wheat</b><br><i>(Triticum </i>L.<i>)</i>	ALMT1 gene region<br>Amplicon length: 95 b.p.	PRF: AATGACTGTGCCGTCTCCAGT<br>PRR: ACAGAGCCGTGTTCTCTGCA<br>PR: ROX-CGTGAAAGCAGCGGAAAGCCTCAGA-BHQ2	Vautrin S. and Zhang D. Real-Time Polymerase Chain Reaction Assay for Endogenous Reference Gene for Specific Detection and Quantification of Common Wheat-derived DNA (Triticum aestivum L.). J. AOAC Int. 2007, 90, 794-801
Species belonging:<br><b>Rapeseed</b><br><i>(Brassica napus)</i>	cruA gene region<br>Amplicon length: 101 b.p.	PRF: GGCCAGGGTTTCCGTGAT<br>PRR: CCGTCGTTGTAGAACCATTGG<br>PR: FAM-AGTCCTTATGTGCTCCACTTTCTGGTGCA-BHQ1	Mazzara M, Grazioli E, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Oilseed Rape Line RT73 Using Real-Time PCR - Validation Report and Protocol. 2007. JRC37550
Species belonging:<br><b>Rice</b><br><i>(Oryza sativa)</i>	PLD gene region<br>Amplicon length: 68 b.p.	PRF: TGGTGAGCGTTTTGCAGTCT<br>PRR: CTGATCCACTAGCAGGAGGTCC<br>PR: ROX-TGTTGTGCTGCCAATGTGGCCTG-BHQ2	Mazzara M, Grazioli E, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Rice Line LLRICE62 Using Real-Time PCR - Validation Report and Protocol. 2006. JRC34091
Species belonging:<br><b>Beet</b><br><i>(Beta vulgaris)</i>	GS2 gene region<br>Amplicon length: 118 b.p.	PRF: GACCTCCATATTACTGAAAGGAAG<br>PRR: GAGTAATTGCTCCATCCTGTTCA<br>PR: ROX-CTACGAAGTTTAAAGTATGTGCCGCTC-BHQ2	Mazzara M, Foti N, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Sugar beet line H7-1 Using Real-Time PCR v. 1.01 - Validation Report and Valideted Method. 2013. JRC84121
Species belonging:<br><b>Soybean</b><br><i>(Glycine max)</i>	le1 gene region<br>Amplicon length: 74 b.p.	PRF: CCAGCTTCGCCGCTTCCTTC<br>PRR: GAAGGCAAGCCCATCTGCAAGCC<br>PR: ROX-CTTCACCTTCTATGCCCCTGACAC-BHQ2	Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011
Species belonging:<br><b>Tomato</b><br><i>(Solanum lycopersicum)</i>	region of tomato pollen protein (LAT52) gene<br>Amplicon length: 89 b.p.	PRF: GACCACGAGAACGATATTTGC<br>PRR: CTTGCCTTTTCATATCCAGACA<br>PR: FAM-CTTTGCAGTCCTCCCTTGGGCT-BHQ1	Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011
Species belonging:<br><b>Cotton</b><br><i>(Gossypium hirsutum)</i>	region of alcohol dehydrogenase C (AdhC) gene<br>Amplicon length: 73 b.p.	PRF: CACATGACTTAGCCCATCTTTGC<br>PRR: CCCACCCTTTTTTGGTTTAGC<br>PR: FAM-TGCAGGTTTTGGTGCCACTGTGAATG-BHQ1	Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011
Species belonging:<br><b>Chicory</b><br><i>(Cichorium </i>L.<i>)</i>	region of actin (ACT) gene<br>Amplicon length: 97 b.p.	PRF: GCCAAATCCAGCTCATCAGT<br>PRR: CCTGTCCGTCGGGTAATTC<br>PR: FAM-ACGAATTACCCGACGGACAGG-BHQ1	GenBit
Species belonging:<br><b>Papaya</b><br><i>(Carica papaya)</i>	region of chymopapain gene<br>Amplicon length: 74 b.p.	PRF: CCATGCGATCCTCCCA<br>PRR: CATCGTAGCCATTGTAACACTAGCTAA<br>PR: FAM-TTCCCTTCAT(BHQ1)CCATTCCCACTCTTGAGA	Nakamura K. et al. Identification and Detection of Genetically Modified Papaya Resistant to Papaya Ringspot Virus Strains in Thailand // Biol. Pharm. Bull. 37(1) 1–5 (2014)

Object

Target

Oligonucleotides

Refernce

Species belonging: Potato (Solanum tuberosum)

region of UDP-glucose pyrophosphorylase gene (UGPase) Amplicon length: 89 b.p.

PRF: GGACATGTGAAGAGACGGAGC PRR: CCTACCTCTACCCCTCCTA PR: ROX-TGAGGAGGTGCGAGGTAATGGTGGTAG-BHQ2

Savini C, Foti N, Mazzara M, Charles Delobel C, Van den Eede G. Event-specific Method for the Quantitation of Event EH92-527-1 Potato Using Real-Time PCR - Validation Report and Protocol. 2006. JRC34758

Species belonging: Maize (Zea mays)

region of zein gene Amplicon length: 82 b.p.

PRF: TTACCGCTTCAGACGATGCC PRR: CATTATTTGCGACACATGCTC PR: FAM-TGACGCCTAACATGATGTCACCA-BHQ1

Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011

Species belonging: Flax (Linum L.)

SAD gene region Amplicon length: 68 b.p.

PRF: GCTCCACCCAGTCACCACCT PRR: TGCGAGGAGATCTGGAGGAG PR: ROX-TGTTGAGGGAGCGTGTTGAAGGGA-BHQ2

Rosario V. et al. Development of an event-specific assay for the qualitative and quantitative detection of the genetically modified flax CDC Triffid (FP967) // Food Control, 2014, 41:128-133

Species belonging: Sunflower (Helianthus annuus)

TUB1 gene region Amplicon length: 128 b.p.

PRF: CGTAGCATATCGTTTCACCG PRR: TCAAGGCAATAGAGATTCCCAA PR: ROX-AGGAAAAATGAGAGAAATCGTAAGCA-BHQ2

GenBit

Species belonging: Wheat (Triticum L.)

ALMT1 gene region Amplicon length: 95 b.p.

PRF: AATGACTGTGCCGTCTCCAGT PRR: ACAGAGCCGTGTTCTCTGCA PR: ROX-CGTGAAAGCAGCGGAAAGCCTCAGA-BHQ2

Vautrin S. and Zhang D. Real-Time Polymerase Chain Reaction Assay for Endogenous Reference Gene for Specific Detection and Quantification of Common Wheat-derived DNA (Triticum aestivum L.). J. AOAC Int. 2007, 90, 794-801

Species belonging: Rapeseed (Brassica napus)

cruA gene region Amplicon length: 101 b.p.

PRF: GGCCAGGGTTTCCGTGAT PRR: CCGTCGTTGTAGAACCATTGG PR: FAM-AGTCCTTATGTGCTCCACTTTCTGGTGCA-BHQ1

Mazzara M, Grazioli E, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Oilseed Rape Line RT73 Using Real-Time PCR - Validation Report and Protocol. 2007. JRC37550

Species belonging: Rice (Oryza sativa)

PLD gene region Amplicon length: 68 b.p.

PRF: TGGTGAGCGTTTTGCAGTCT PRR: CTGATCCACTAGCAGGAGGTCC PR: ROX-TGTTGTGCTGCCAATGTGGCCTG-BHQ2

Mazzara M, Grazioli E, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Rice Line LLRICE62 Using Real-Time PCR - Validation Report and Protocol. 2006. JRC34091

Species belonging: Beet (Beta vulgaris)

GS2 gene region Amplicon length: 118 b.p.

PRF: GACCTCCATATTACTGAAAGGAAG PRR: GAGTAATTGCTCCATCCTGTTCA PR: ROX-CTACGAAGTTTAAAGTATGTGCCGCTC-BHQ2

Mazzara M, Foti N, Savini C, Van den Eede G. Event-specific Method for the Quantitation of Sugar beet line H7-1 Using Real-Time PCR v. 1.01 - Validation Report and Valideted Method. 2013. JRC84121

Species belonging: Soybean (Glycine max)

le1 gene region Amplicon length: 74 b.p.

PRF: CCAGCTTCGCCGCTTCCTTC PRR: GAAGGCAAGCCCATCTGCAAGCC PR: ROX-CTTCACCTTCTATGCCCCTGACAC-BHQ2

Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011

Species belonging: Tomato (Solanum lycopersicum)

region of tomato pollen protein (LAT52) gene Amplicon length: 89 b.p.

PRF: GACCACGAGAACGATATTTGC PRR: CTTGCCTTTTCATATCCAGACA PR: FAM-CTTTGCAGTCCTCCCTTGGGCT-BHQ1

Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011

Species belonging: Cotton (Gossypium hirsutum)

region of alcohol dehydrogenase C (AdhC) gene Amplicon length: 73 b.p.

PRF: CACATGACTTAGCCCATCTTTGC PRR: CCCACCCTTTTTTGGTTTAGC PR: FAM-TGCAGGTTTTGGTGCCACTGTGAATG-BHQ1

Van den Eede G. Compendium of Reference Methods for GMO Analysis. JRC Reference Reports. 2011

Species belonging: Chicory (Cichorium L.)

region of actin (ACT) gene Amplicon length: 97 b.p.

PRF: GCCAAATCCAGCTCATCAGT PRR: CCTGTCCGTCGGGTAATTC PR: FAM-ACGAATTACCCGACGGACAGG-BHQ1

GenBit

Species belonging: Papaya (Carica papaya)

region of chymopapain gene Amplicon length: 74 b.p.

PRF: CCATGCGATCCTCCCA PRR: CATCGTAGCCATTGTAACACTAGCTAA PR: FAM-TTCCCTTCAT(BHQ1)CCATTCCCACTCTTGAGA

Nakamura K. et al. Identification and Detection of Genetically Modified Papaya Resistant to Papaya Ringspot Virus Strains in Thailand // Biol. Pharm. Bull. 37(1) 1–5 (2014)

Performance of the split "cp4 epsps" system

Line	cp4 epsps1	cp4 epsps2	cp4 epsps1 + cp4 epsps2
GTS 40-3-2	+	-	-
MON89788	-	+	-
NK603	+	+	-
MON88017	+	+	-
RT73 (GT73)	-	+	-
H7-1	-	+	-
GTS-B77	-	+	-