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Certification of a reference material of Campylobacter coli and jejuni (CNET068 and CNET112) agarose plugs for PFGE: IRMM-313

Abstract

This report describes the production of IRMM-313, a Campylobacter coli and jenuni gDNA material certified for the size of the DNA fragments obtained by enzymatic restriction and Pulsed Field Gel Electrophoresis (PFGE). The material was produced following ISO Guide 34:2009. The CRM was produced from cultures of Campylobacter coli CNET068 and jejuni CNET112 which were pooled and processed into agarose plugs suited for PFGE. The bacterial cells were lysed as to release the gDNA within the plug. Between unit-homogeneity and stability during dispatch and storage were assessed in accordance with ISO Guide 35:2006. The material was characterised by an intercomparison among laboratories of demonstrated competence and complying to ISO/IEC 17025. Technically invalid results were removed but no outlier was eliminated on statistical grounds only. The certified values were obtained by PFGE. Uncertainties of the certified values were calculated in compliance with the Guide to the Expression of Uncertainty in Measurement (GUM). The material is intended for quality control and assessment of method performance. As any reference material, it can also be used for control charts or validation studies. The CRM is available in plastic screw cap vials containing 1 plug suspended in Tris EDTA buffer solution. The minimum amount of sample recommended to be used is ½ plug.JRC.D.2-Standards for Innovation and sustainable Developmen

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