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ID   QL-CON-00-015; SV 0; linear; genomic DNA; STS; SYN; 136 BP.
XX
AC   ;
XX
DT   16-JUL-2013
DT   18-OCT-2021
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DE   Qualitative PCR method for detection of the junction between the CaMV35S promoter and the bar gene (verified by the EURL GMFF in the context of Commission Decision 2006/578/EC).
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KW   construct_specific, EURL_GMFF_in-house_verified.
XX
OS   Oryza sativa (rice).
XX
RN   [1]
RP   1-136
RA   Bayer CropScience;
RT   "Grain testing method for detection of rice GM event LLRICE601 using RT-PCR protocols PGS0505 and PGS0476";
RL   Online Publication (2006).
RX   CRL-DOC=P35S-bar sqRT-PCR - PGS0494-476 310806.pdf
XX
RN   [2]
RP   1-136
RA   US Department of Agriculture;
RT   "Verification of the Bayer CropScience method for the detection of LL601 in rice using real-time PCR";
RL   Online Publication (2006).
RX   CRL-DOC=35SBarRiceVerficationReportrev.pdf
XX
RN   [3]
RP   1-136
RA   European Union Reference Laboratory for GM Food and Feed (EURL GMFF), Joint Research Centre (JRC), European Commission;
RT   "Report on the Verification of a Construct-specific Detection Method for
RT   Identification of Rice GM-Events containing P35S::BAR using a Real-time
RT   PCR Assay";
RL   Online Publication (2006).
RX   CRL-DOC=Verification Report 35S-BAR CRL.pdf
XX
RN   [4]
RP   1-136
RT   "PCR reactions set up and amplification conditions";
RL   Online Publication (2006).
RX   PCR=QL-CON-00-015.pdf
XX
DR   GMOMETHODS; QL-TAX-OS-002;
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FH   Key             Location/Qualifiers
FH
FT   STS             1..136
FT                   /standard_name="PCR 136 bp amplicon"
FT                   /note="Construct-specific RT-PCR"
FT                   /target="Junction region between the Cauliflower Mosaic Virus 35S promoter (CaMV P-35S) and the phosphinothricin N-acetyl transferase (bar) gene from Streptomyces hygroscopicus"
FT   misc_feature    1..136
FT                   /note="The fragment size generated by the CaMV P-35S-bar PCR depends on the assembly of the two sequence elements and is varying in the different GM plant events"
FT   primer_bind     1..21
FT                   /standard_name="Primer forward: MDB498"
FT                   /note="TATCCTTCGCAAGACCCTTCC"
FT                   /target="CaMV P-35S"
FT   primer_bind     22..46
FT                   /standard_name="RT-PCR probe: TM099"
FT                   /note="FAM-TCTATATAAGGAAGTTCATTTCATT-MGBNFQ"
FT   primer_bind     complement(115..136)
FT                   /standard_name="Primer reverse: DPA143"
FT                   /note="ATGTCGGCCGGGCGTCGTTCTG"
FT                   /target="bar gene"
XX
SQ   Sequence 136 BP; 37 A; 41 C; 21 G; 37 T; 0 other;
     tatccttcgc aagacccttc ctctatataa ggaagttcat ttcattnnnn nnnnnnnnnn        60
     nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnncagaac       120
     gacgcccggc cgacat                                                       136
//