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Detection Image Detection method details

Name:

 QL-CON-00-015

Description:

 Qualitative real-time PCR (TaqMan) method for detection of the junction between the CaMV 35S promoter and the bar gene (Bayer CropScience, 2006; verified by the EU-RL GMFF in the context of Commission Decision 2006/578/EC)

Comment:

 Target is the junction between the Cauliflower mosaic virus 35S promoter (CaMV P-35S) and the phosphinothricin N-acetyltransferase (bar) gene from Streptomyces hygroscopicus.

The fragment size generated by the QL-CON-00-015 PCR depends on the assembly of the two sequence elements and is varying in the different GM plant events. The sequence data for the Verdeca HB4 soybean and CM8101 maize events, for example, indicates that the L-TEV element is inserted between P-35S and bar, so a PCR amplicon of 236 bp should be expected.

Validation:

 in-house validation

Standardisation:

 none

Related Methods:

Type:

 construct-specific

Target GMO names:
CBH351, CM8101, DBT418, DLL25, EH-2 modbs 2.99, IE034, Iksan515, Iksan526, LLCotton25, LLRICE06, LLRICE601, LLRICE604, LLRICE62, MON88701, Ms3, Ms6, ND207, SYHT04R, T1c-19, T2A-1, T303-3, T304-40, TC6275, Varuna bn 3.6, Verdeca HB4, W62, W98

  • Oligonucleotides:

  • Forward Primer

  • Name:

    MDB498

  • Sequence:

    TATCCTTCGCAAGACCCTTCC

  • Size:

    21

  • Reverse Primer

  • Name:

    DPA143

  • Sequence:

    ATGTCGGCCGGGCGTCGTTCTG

  • Size:

    22

  • Probe

  • Name:

    TM099

  • Sequence:

    TCTATATAAGGAAGTTCATTTCATT

  • Size:

    25

  • Amplicon:

  • Sequence:

    tatccttcgcaagacccttcctctatataaggaagttcatttcatttggagaggacagggtaccggatccaacaatgagcccagaacgacgcccggccgacat

  • Size:

    103

Documents

Citation Type Local copy
Grain testing method for the detection of rice GM events containing 35S::bar sequences using RT-PCR protocols PGS0494 and PGS0476. (2006) Bayer CropScience. BioScience 31-8-2006, Ghent, BelgiumLink to the document url.
EU-RL GMFF GMOMETHODS database; QL-CON-00-015Link to the document url. EU reference method protocol