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Detection Image Detection method details

Name:

 QL-CON-00-011

Description:

 Qualitative real-time PCR (TaqMan) method for detection of the junction between the 35S promoter and the phosphinothricin N-acetyltransferase (pat) gene (ISO 21569-3).

Comment:

 Target is the junction region between the 35S promoter (P-35S) from the Cauliflower mosaic virus and a modified phosphinothricin N-acetyltransferase (pat) gene from Streptomyces viridochromogenes (the amplicon size may vary depending on the P-35S/pat construct present in the event, e.g. in DAS59122 it is 102 bp long).

Validation:

 ring trial validation

Standardisation:

 ISO standard

Related Methods:

Type:

 construct-specific

Target GMO names:
32316, 40416, 4114, 43A47, 676, 678, 680, A2704-12, A2704-21, A5547-127, A5547-35, Bt10 maize, Bt11, DAS1507, DAS59122, DAS59132, DBN9004, DHA, Falcon GS 40/90, GU262, HCN10, HCR-1, Liberator, MZHG0JG, MZIR098, SYHT0H2, T120-7, T14, T25, T45, Topas 19/2

  • Oligonucleotides:

  • Forward Primer

  • Name:

    35SP03.f

  • Sequence:

    AAGTTCATTTCATTTGGAGAGGACA

  • Size:

    25

  • Reverse Primer

  • Name:

    pat-7.r

  • Sequence:

    CGGCCATATCAGCTGCTGTAG

  • Size:

    21

  • Probe

  • Name:

    GSS01.s

  • Sequence:

    CCGGAGAGGAGACCAGTTGAGATTAGGC

  • Size:

    28

  • Amplicon:

  • Sequence:

    aagttcatttcatttggagaggacannnnnnnnnnnnnnnnnnnnnnnnnccggagaggagaccagttgagattaggcnnnctacagcagctgatatggccg

  • Size:

    102

Documents

Citation Type Local copy
Methodensammlung des Länderausschusses Gentechnik. Real-Time PCR zur quantitativen Bestimmung gentechnisch veränderter Rapslinien mit dem 35S/pat-Genkonstrukt. Erstellt vom Unterausschuss Methodenentwicklung; Stand 24.03.2006Link to the document url. other document
Horizontal methods for molecular biomarker analysis — Methods of analysis for the detection of genetically modified organisms and derived products — Part 3: Construct-specific real-time PCR method for detection of P35S-pat-sequence for screening for genetically modified organisms (ISO/TS 21569-3)Link to the document url. other document
EU-RL GMFF GMOMETHODS database; QL-CON-00-011Link to the document url. EU reference method protocol
Real-Time PCR zur quantitativen Bestimmung gentechnisch veränderter Rapslinien mit dem 35S/pat-GenkonstruktLink to the document url. EU reference method protocol