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Detection Image Detection method details

Name:

QL-ELE-00-019

Description:

Qualitative real-time PCR (SYBRGreen) method for detection of CS-CP4epsps-RHIRD (Barbau-Piednoir et al., 2014)

Comment:

Target is the CS-CP4epsps-RHIRD from Agrobacterium tumefaciens

Two reverse primers are described that are meant to be used in PCR together. Primer CP4 Synthetic R (TGAAGGACCGGTGGGAGAT) binds to the nucleotide sequence of CS-CP4epsps-RHIRD in soybean, maize and cotton GMO events whereas primer CP4 synth Rbis (TGAAGGACCTGTGGGAGAT) recognizes a nucleotide sequence of CS-CP4epsps-RHIRD in GM rapeseed and sugar beet (Barbau-Piednoir et al., 2012). For Bioinformatical reasons, a degenerate primer was introduced in the reverse primer sequence (K=G/T) and in the amplicon.

Validation:

ring trial validation

Standardisation:

unknown

Related Methods:

QL-TAX-BN-003, QL-TAX-GM-001, QL-TAX-ZM-002

Type:

element-specific

Target DNA element:

CS-CP4epsps-RHIRD
  • Oligonucleotides:

  • Forward Primer

  • Name:

    CP4 Synthetic F
  • Sequence:

    GCATGCTTCACGGTGCAA
  • Size:

    18
  • Reverse Primer

  • Name:

    CP4 Synthetic R / CP4 synth Rbis
  • Sequence:

    TGAAGGACCKGTGGGAGAT
  • Size:

    19
  • Amplicon:

  • Sequence:

    gcatgcttcacggtgcaannnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnnatctcccacmggtccttca
  • Size:

    108

Documents

Citation Type Local copy
Barbau-Piednoir, E., Stragier, P., Roosens, N., Mazzara, M., Savini, C., Van den Eede, G., Van den Bulcke, M. (2014). Inter-laboratory testing of GMO detection by combinatory SYBR®Green PCR screening (CoSYPS). Food Analytical Methods. 7, 1719-1728.Link to the document url. peer-reviewed article
EU-RL GMFF GMOMETHODS database; QL-ELE-00-019Link to the document url. EU reference method protocol
Barbau-Piednoir, E. et al. (2012). Four new SYBR_Green qPCR screening methods for the detection of Roundup Ready, LibertyLink, and CryIAb traits in genetically modified products. Eur Food Res Technol, 234, 13–23.Link to the document url. peer-reviewed article 09-01-2015