# Commission Regulation (EU) No 175/2010 of 2 March 2010 implementing Council Directive 2006/88/EC as regards measures to control increased mortality in oysters of the species Crassostrea gigas in connection with the detection of Ostreid herpesvirus 1 μvar (OsHV-1 μvar) (Text with EEA relevance), Division 3: Equipment and environmental conditions

Extent: E+W+S+N.I.

The diagnostic test used for OsHV-1 μvar detection and identification by PCR requires the equipment and environmental conditions classically used for PCR assays as follows:
A closed hood equipped with an UV producing system to eliminate potential contaminations when preparing PCR mix.
Two complete sets of pipettes (2 μl; 20 μl; 200 μl and 1 000 μl), the first one for DNA extraction, and the second one for PCR mix preparation.
Three different pipettes: one pipette (2 μl) to dispense samples in PCR mix, one pipette (20 μl) for EB sampling and another pipette (20 μl) to load PCR products in agarose gels.
Filter pipette tips (2 μl; 20 μl; 200 μl and 1 000 μl) for DNA extraction, PCR mix preparation and sample dispensing.
Pipette tips (20 μl) to collect EB and to load amplification products in agarose gel.
A thermal cycler to perform amplifications.
A horizontal electrophoresis system for PCR products electrophoresis.
An UV table to observe PCR products after agarose gel electrophoresis.
A system to acquire pictures of the gels.
The manipulator must wear a lab coat and some gloves during all the different steps described bellow. Lab coat and gloves must be changed preferably after each main step: DNA extraction, preparation of PCR mix, sample dispensing, amplification and gel loading.
It is recommended to perform these different steps in different rooms. More particularly, amplification and gel loading/electrophoresis should take place in a room separate from DNA extraction, PCR mix preparation and DNA dispensing.

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Source: https://www.legislation.gov.uk/eur/2010/175/annex/I/part/B/division/3 (the official version)
Page: https://openlegislation.uk/eu-regulation-2010-175/annex-I/part-B/division-3
Contains public sector information licensed under the Open Government Licence v3.0. Source: legislation.gov.uk, The National Archives.