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European Union Reference Laboratory for Genetically Modified Food and Feed (EURL GMFF)


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ID   QT-CON-00-007; SV 0; linear; genomic DNA; STS; SYN; 100 BP.
XX
AC   SYN-EV176-9;
XX
DT   15-SEP-2009
DT   05-OCT-2016
XX
DE   Quantitative PCR method for detection of the junction between a synthetic cry1A(b) gene and the Phospho-enol-pyruvate Carboxylase intron N.9.
XX
KW   construct_specific.
XX
OS   Zea mays (maize) - event Bt176 (SYN-EV176-9)
XX
RN   [1]
RP   1-100
RA   Shindo Y., Kuribara H., Matsuoka T., Futo S., Sawada C., Shono J.,
RA   Akiyama H., Goda Y., Toyoda M., Hino A.;
RT   "Validation of real-time PCR analyses for line-specific quantitation of
RT   genetically modified maize and soybean using new reference molecules";
RL   J AOAC Int 85:1119-1126 (2002).
RX   PUBMED; 12374412.
XX
RN   [2]
RP   1-100
RT   "Foodstuffs - Methods of analysis for the detection of genetically
RT   modified organisms and derived products - Quantitative nucleic acid based
RT   methods";
RL   ISO 21570:1-103 (2005).
RX   ISO=34615
XX
RN   [3]
RP   1-100
RT   "PCR reactions set up and amplification conditions";
RL   Online Publication (2010).
RX   PCR=QT-CON-00-007.pdf
XX
DR   GMOMETHODS; QT-TAX-ZM-006;
XX
FH   Key             Location/Qualifiers
FH
FT   STS             1..100
FT                   /standard_name="PCR 100 bp amplicon"
FT                   /note="Construct-specific RT-PCR"
FT                   /target="Junction region between a synthetic cry1A(b) gene and the Phospho-enol-pyruvate Carboxylase (PEPC) intron N.9"
FT   primer_bind     1..20
FT                   /standard_name="Primer forward: E176 2-5'"
FT                   /note="TGTTCACCAGCAGCAACCAG"
FT                   /target="cry1A(b)"
FT   primer_bind     21..75
FT                   /standard_name="RT-PCR probe: E176-Taq"
FT                   /note="FAM-CCGACGTGACCGACTACCACATCGA-TAMRA"
FT   primer_bind     complement(76..100)
FT                   /standard_name="Primer reverse: E176 2-3'"
FT                   /note="ACTCCACTTTGTGCAGAACAGATCT"
FT                   /target="IVS 9 PEPC"
XX
SQ   Sequence 100 BP; 13 A; 11 C; 11 G; 10 T; 55 other;
     tgttcaccag cagcaaccag nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn        60
     nnnnnnnnnn nnnnnagatc tgttctgcac aaagtggagt                             100
//