Community Research and Development Information Service - CORDIS

Abstract

This report describes the production and certification of a reference material consisting of purified plasmid DNA (pDNA) pUC18 containing a 609 base pairs fragment of human Factor II (prothrombin) gene sequence, IRMM/IFCC-490. This certified reference material (CRM) is intended to support the validation and the harmonisation of polymerase chain reaction (PCR)-based methods used for detection of the G20210A mutation in the human prothrombin gene. The homogeneity and short-term stability of the batch was assessed by monitoring the melting temperature (Tm) of the product and the crossing point threshold (Cp) value obtained after PCR amplification using the LightCycler Factor II (prothrombin) G20210A kit and a LightCycler 1.5 instrument. The long-term stability was assessed using the cycle threshold value (Ct) obtained after real-time PCR amplification using specific primers and probe in combination with ABI Prism instruments.
The material is homogeneous and stable at -20 °C. The sequence of the insert was confirmed by DNA sequence analysis. Each vial contains an indicative volume of 50 µL corresponding to approximately 1 ng of pDNA.

Additional information

Authors: GANCBERG D, European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Geel (BE);MARKI-ZAY J, European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Geel (BE);CORBUSIER P, European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Geel (BE);SCHIMMEL H, European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Geel (BE);EMONS H, European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Geel (BE);C.KLEIN, European Commission, Joint Research Centre, Institute for Health and Consumer Protection, Ispra (IT)
Bibliographic Reference: EUR 22169 EN (2006),35 pp. Free of charge
Availability: Available from European Commission, JRC Knowledge Management Unit, Ispra (IT) Tel: +39 033278 9893 Fax: +39 033278 5409 E-mail: jrc-publications-office@ec.europa.eu or Bruxelles (BE) Tel: +32 2 295 76 24 Fax: +32 2 299 63 22
ISBN: ISBN: 92-79-01633-4
Record Number: 200618677 / Last updated on: 2006-08-11
Category: PUBLICATION
Original language: en
Available languages: en