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Abstract

This report describes the production and certification of IRMM-312, a reference material of genomic DNA (gDNA) of Bacillus subtilis DSM 5750 in agarose inserts. This CRM (IRMM-312) is intended to be used for the taxonomic identification of authorised probiotic feed additives by Pulsed Field Gel Electrophoresis (PFGE). The homogeneity and stability of the batch was assessed by monitoring the PFGE pattern of a SfiI restriction enzyme digest of gDNA of Bacillus subtilis DSM 5750 in agarose inserts. The batch was found to be homogeneous and the material is stable at 4 °C. The batch was characterised by five laboratories determining the PFGE pattern of a SfiI restriction enzyme digest of IRMM-312. The pattern should be considered as a whole and restriction fragments in the size range from 15 kb to 97 kb are certified for their fragment length. DNA sequence analysis of the 3' end of the 16S rRNA gene and the entire 16S-23S internal transcribed spacer (ITS) coding region confirmed the identity of the strain as Bacillus subtilis. Each vial contains one agarose insert containing gDNA of Bacillus subtilis DSM 5750.

Additional information

Authors: DE BAETS L, European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Geel (BE);MEEUS N, European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Geel (BE);VAN IWAARDEN P, European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Geel (BE);PHILIPP W, 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)
Bibliographic Reference: EUR 23372 EN (2008), 29 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-info@ec.europa.eu or Bruxelles (BE) Tel: +32 2 295 76 24 Fax: +32 2 299 63 22
ISBN: ISBN: 978-92-79-09026-4
Record Number: 200819854 / Last updated on: 2008-06-10
Category: PUBLICATION
Original language: en
Available languages: en
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