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Detection Image Detection method details

Name:

 QL-ELE-00-024

Description:

 Qualitative real-time PCR (TaqMan) method for detection of the terminator of the pea ribulose-1,5-bisphosphate carboxylase small subunit (rbcS) E9 gene (EU-RL GMFF GMOMETHODS database as of January 20, 2017; Official Collection of Methods according to § 28b GenTG, G 30.40-22).

Comment:

 Target is the E9 (T-E9) terminator from pea (Pisum sativum).

As described in G 30.40-22, the PCR was optimised by modifying the chemistry (in particular, by introducing the use of an internal quencher in the probe).

Validation:

 ring trial validation

Standardisation:

 EU reference method

Related Methods:

Type:

 element-specific

Target DNA element:
T-rbcS_E9-PEA

  • Oligonucleotides:

  • Forward Primer

  • Name:

    T-E9-F

  • Sequence:

    TGAGAATGAACAAAAGGACCATATCA

  • Size:

    26

  • Reverse Primer

  • Name:

    T-E9-R

  • Sequence:

    TTTTTATTCGGTTTTCGCTATCG

  • Size:

    23

  • Probe

  • Name:

    T-E9-P

  • Sequence:

    TCATTAACTCTTCTCCATCCATTTCCATTTCACAGT

  • Size:

    36

  • Amplicon:

  • Sequence:

    tgagaatgaacaaaaggaccatatcattcattaactcttctccatccatttccatttcacagttcgatagcgaaaaccgaataaaaa

  • Size:

    87

Documents

Citation Type Local copy
Debode, F., Janssen, E., Berben, G. (2013). 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, 236, 659–669.Link to the document url. peer-reviewed article
Detection of T-E9 and T-g7 terminator sequences for screening genetically modified plants using real-time PCR (singleplex)Link to the document url. other document