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Screening of Beer-Spoilage Bacteria Using,,,the LightCycler PCR Workflow System

rification quality by Magnetic Bead Technology
  • Improved reproducibility by precise pipetting steps

  • Sample names were directly imported with a bar-code reader and transferred via network from the MagNA Pure Instrument to the LightCycler Instrument. This minimized the risk of wrong entries and further reduced the hands-on time. Utilizing the MagNA Pure Total NA Large Volume protocol meant that the enriched samples could be directly used for nucleic acid purification without any additional centrifugation steps. For sample preparation, 1 ml of enrichment culture was predispensed into the wells of the MagNA Pure Sample Cartridge. Following fully automated nucleic acid purification, DNA was eluted in 50 l final volume. The complete PCR mixes were then set up automatically. The MagNA Pure LC Instrument compiled and loaded the PCR mixes directly into LightCycler Capillaries in the LightCycler Carousel.

    PCR setup

    For one reaction of 20 l final volume, 13 l LightCycler foodproof Beer Screening Mastermix, 1 l LightCycler foodproof Beer Screening Enzyme Solution and 1 l LightCycler foodproof Beer Screening Internal Control were automatically mixed and pipetted together with 5 l sample DNA into each LightCycler Capillary.

    Suitability testing of routine samples

    58 routine samples including control, different kinds of beer, fermenter, rinsing water and other yeast-containing samples were tested. To assess the efficiency of the MagNA Pure LC System in removing PCR inhibitors, the purified DNA was analyzed with the LightCycler foodproof Beer Screening Kit.

    Sensitivity and specificity testing of dilution series

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