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Method of reference and SOPs

Horse DNA detection method

 

Method of reference for the detection of animal proteins in feed

The Annex VI of Commission Regulation n° 152/2009 was entirely revised and amended by Commission Regulation n° 51/2013 published on the 16th of January 2013

The new regulation relies on the combination of light microscopy and real time PCR for the detection of animal proteins in feed in the European Union. This regulation also relies on Standard Operating Procedures (SOPs) edited by the EURL-AP for technical implementation. These SOPs are binding or optional complements to the regulation and are available for download on this page.

 

Calibration kits for ruminant and pig detection by PCR

The calibration kits ERM-AD482 RUMINANT pDNA CALIBRANT and ERM-AD483 PORCINE pDNA CALIBRANT can be ordered from JRC Geel. They are now Certified European Reference Materials.

For ordering and information, go to the following webpages :
https://crm.jrc.ec.europa.eu/p/erm-AD482
https://crm.jrc.ec.europa.eu/p/erm-AD483

 

List of approved master mixes

Detection of ruminant DNA

As mentioned in the  Commission Regulation (EU) No 51/2013 as well as in the SOP “ Detection of ruminant DNA in feed using real-time PCR”, it is crucial to be careful that the master mix used is fit for the purpose and does not contain bovine serum albumin (BSA) or other reagents susceptible to lead to false results due to presence of animal DNA.

The EURL-AP recommends to use one of the following mastermixes:

  • “Universal mastermix” reference number DMML-D2-D600 (Diagenode, Liège, Belgium) www.diagenode.com
  • “qPCR MasterMix” reference number RT-QP2X-03 (Eurogentec, Seraing, Belgium) www.eurogentec.com   

 

List of SOPs for download

This list is updated and contains only valid versions. The binding or optional character of a SOP is indicated.  The original version is labelled V.1.0. Minor revisions are labelled V.1.# and major revisions are labelled V.#.0. SOPs recently uploaded are tagged NEW for two months after publication date.

Title

Version

   

EURL-AP SOP slide preparation and mounting

V.1.0

BINDING


EURL-AP SOP use of staining reagents

V.1.0

OPTIONAL


EURL-AP SOP DNA extraction

V.1.1

BINDING

 

EURL-AP SOP operational schemes

V.3.0 BINDING  

EURL-AP SOP Ruminant PCR

V.1.2 BINDING NEW

EURL-AP File for cut-off determination exact copy number + copies cut-off

V.2.0 BINDING