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mericon DNA Bacteria Kit and mericon DNA Bacteria Plus Kit

For extraction of high-quality DNA from microbial enrichment culture

  • Fast and efficient lysis of both Gram-negative and Gram-positive bacteria
  • Protocols based on mechanical and thermal lysis
  • Efficient removal of PCR inhibitors
The mericon DNA Bacteria Kit and mericon DNA Bacteria Plus Kit are designed to enable extraction of high-quality nucleic acids from Gram-positive and Gram-negative bacteria. The kits form part of the comprehensive QIAGEN food testing portfolio, which also features assays for pathogen and GMO DNA detection and ingredient authentication.
Cat No./ID: 69525
mericon DNA Bacteria Kit (100)
€124.00
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Fast Lysis Buffer
Cat No./ID: 69534
mericon DNA Bacteria Plus Kit (50)
€124.00
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50 Pathogen Lysis Tubes L, Fast Lysis Buffer
The mericon DNA Bacteria Kit and mericon DNA Bacteria Plus Kits are intended for molecular biology applications in food, animal feed, and pharmaceutical product testing. This product is not intended for the diagnosis, prevention, or treatment of a disease.

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Efficient DNA extraction from Gram-negative and Gram-positive bacteria.

(A) Samples of an enrichment culture of Listeria monocytogenes were diluted and subjected to the mechanical (bead) lysis protocol of the mericon DNA Bacteria Plus Kit and the thermal (boiling) lysis protocol of the mericon DNA Bacteria Kit to evaluate lysis efficiency. For this Gram-positive bacterium, mechanical lysis delivered a more effective extraction, most evident at very low bacterial concentrations. (B) Diluted and undiluted samples of an enrichment culture of Salmonella enterica in milk and hazelnut spread were subjected to the mechanical and thermal lysis protocols. Overall, the efficiency of DNA extraction from Gram-negative bacteria is similar with both preparation protocols.
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Highly sensitive pathogen detection.

Detection of pathogens was highly sensitive, even in difficult food matrices, such as peanut butter. Peanut butter was homogenized in buffered peptone water, spiked with <5 cfu of salmonella, and enriched for 20 h at 37°C. DNA was extracted from serially diluted samples of the enrichment culture using the mericon DNA Bacteria Kit and then tested with the mericon Salmonella Kit. Although the original inoculation was small, salmonella was still reliably detected at a dilution factor of 1:100,000.
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Comparison of time to result of different methods for pathogen detection.

Delivering sensitive and highly specific results, real-time PCR analysis is also faster than conventional pathogen detection methods. Besides taking longer, immunoassays and traditional culture methods are less sensitive and more labor intensive.

Performance
The thermal lysis procedure using the mericon DNA Bacteria Kit is optimized for the majority of easily lysed Gram-negative bacteria (e.g., Salmonella spp., Camplylobacter spp., and Cronobacter spp.), whereas the advanced features of the mechanical sample preparation using the mericon DNA Bacteria Plus Kit create the lysis conditions needed for difficult Gram-positive bacteria, such as Listeria spp. (see Figure "Efficient DNA extraction from Gram-negative or Gram-positive bacteria").
Principle
To date, the predominant method to test food for the presence of pathogens has been via culture preparation and/or ELISA-based assays. Molecular-based methods, specifically real-time PCR, offer an informative complement to these well-established methods. In addition, real-time PCR detection delivers results in less time (see Figure "Comparison of time to result of different methods for pathogen detection"), affords greater sensitivity and specificity, and has the robustness to cope with difficult food matrices (see Figure "Highly sensitive pathogen detection"). Finally, real-time PCR is a safe method for detecting challenging pathogen strains.

Procedure

To provide a streamlined and straightforward method for microbial sample preparation from pathogen enrichment cultures, QIAGEN has designed protocols based on mechanical lysis (using beads) and thermal lysis (by boiling) that deliver optimal disruption force for different bacteria types.

Thermal lysis is highly suited for the majority of easily lysed Gram-negative bacteria, whereas mechanical sample preparation creates the lysis conditions needed for difficult-to-lyse Gram-positive bacteria.

Carefully formulated chemistry included in the mericon DNA Bacteria Kit and the mericon DNA Bacteria Plus Kit maximizes lysis efficiency, stabilizes the extracted DNA, and removes inhibitors that may interfere with downstream applications.

Applications
The mericon DNA Bacteria and mericon DNA Bacteria Plus Kits are designed for the extraction of total genomic DNA from Gram-negative bacteria (mericon DNA Bacteria Kit) and Gram-positive bacteria (mericon DNA Bacteria Plus Kit) from pathogen enrichment cultures.
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