From Bulk Milk and Blood with Vacuum Technology

The aim of this research application was to improve molecular methods for the detection of bovine viral diarrhoea virus (BVDV) with the expectation of applying the knowledge learned to advance the study of the related classical swine fever virus.

Total RNA is extracted either from bulk milk or whole anti-coagulated blood on the MICROLAB STAR using the Promega SV96 Total RNA Kit protocol, omitting the DNase step. The extraction takes approximately 90 minutes to process 96 samples starting from lysis and ending with eluted RNA. After RNA isolation, the presence or absence of the specific virus RNA is checked with a series of specific PCR reactions, which are analysed on gels. The PCR reaction set-up under mineral oil is fully automated on the system, and the following analysis of the PCR products on gels is done manually.

Using the MICROLAB STAR for automated RNA isolation and PCR set-up during these studies, the very first type 2 bovine viral diarrhoea virus was isolated and characterized in the UK (see publications).

Graham S. Heath, Donald P. King, Jane L.E. Turner, Philip R. Wakeley, Malcolm Banks. 2003. Use of an internal standard in a TaqMan nested reverse transcription-polymerase chain reaction for the detection of bovine viral diarrhoea virus. Vet. Microbiology. 96, 357-366.

P.R. Wakeley, J.L.E. Turner, G. Ibata, D.P. King, T Sandvik, P. Howard, T.W. Drew. 2004. Characterization of a type 2 bovine viral diarrhoea virus from the UK. Vet. Microbiology. 102, 19-24.

Features and
Benefits

  • More consistent total RNA yield and quality from field samples in comparison to manual processing
  • Full automation of RNA isolation and PCR set-up increases throughput
  • Error-free pipetting for reliable processing of the samples

System
Configuration

  • MICORLAB STAR, 4 channels, Autoload option with barcode reader
  • Qiagen QIAvac 96 Manifold Assembly, mounted on special carrier
  • Actively-cooled carrier for PCR plate
  • Thermo Haake DC10 Circulation Bath
  • Vacutainer Tube Carriers

Links

Links

www.promega.com
(kits)

 www.qiagen.com
(QAIvac)

 www.thermohaake.com
(circulation bath)

 
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