For extraction of high-quality DNA from microbial enrichment cultures and raw or processed foods

  • Modified CTAB procedure ensures faster sample processing
  • Efficient isolation of even fragmented DNA from processed foods
  • Fast and efficient lysis of Gram-negative and Gram-positive bacteria
  • No carryover of PCR inhibitors from complex food matrices
mericon Sample Preparation Kits are designed to enable the extraction of high-quality nucleic acids from a wide range of starting materials, such as pathogen enrichment cultures (mericon DNA Bacteria and Bacteria Plus Kits) and a variety of raw and processed foods (DNeasy mericon Food Kit). The carryover of PCR inhibitors from these complex food matrices is minimized. The isolated, pure DNA is suitable for downstream applications, such as real-time PCR ingredient authentication, pathogen detection, or GMO DNA detection and quantification using mericon PCR Assay Kits.
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Produto Num Cat. Preço de lista:
DNeasy mericon Food Kit (50)
50 QIAquick Spin Columns, Proteinase K, buffers
69514
Cotação
mericon DNA Bacteria Kit (100)
Fast Lysis Buffer
69525
Cotação
mericon DNA Bacteria Plus Kit (50)
50 Pathogen Lysis Tubes L, Fast Lysis Buffer
69534
Cotação
mericon Sample Preparation Kits are intended for molecular biology applications in food, animal feed, and pharmaceutical product testing. These products are not intended for the diagnosis, prevention, or treatment of a disease.
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High yields of DNA with minimal inhibitor carryover.|Efficient bacterial DNA extraction.|Efficient and rapid DNA extraction.|mericon DNA Bacteria Kit.|mericon DNA Bacteria Plus Kit.|
[A] Samples of peanut butter and chocolate were prepared using the DNeasy mericon Food Kit (standard protocol) and products from 3 other suppliers to amplify a segment of the tnrL chloroplast tRNA gene. The DNeasy mericon Food Kit yielded a significantly lower CT value than kits from the other suppliers, indicating a higher yield of DNA. *Only one replicate resulted in DNA amplification. [B] To test for inhibitors carried over from the sample preparation procedure, samples were diluted 1:10 and the same DNA segment was amplified. Both food samples were inhibitor-free when prepared with the DNeasy mericon Food Kit. Samples prepared with products from the other suppliers showed strong inhibitor contamination.|[A] Samples of an enrichment culture of Listeria monocytogenes were diluted and subjected to the mechanical lysis protocol of the mericon DNA Bacteria Plus Kit and the thermal 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 was similar with both preparation protocols.|The DNeasy mericon Food Kit enables DNA extraction from a whole range of food samples in a significantly shorter time than conventional CTAB protocols.|||
Desempenho
DNeasy mericon Food Kit

During production, food is subjected to heat, irradiation, high pressure, and changes in pH which lead to DNA degradation and fragmentation. The optimized chemistry of the DNeasy mericon Food Kit was developed to recover short DNA fragments (down to 100 bp) ensuring that even highly fragmented DNA is efficiently isolated and subsequently amplified in PCR reactions. Thanks to these features, this kit offers a universally applicable extraction method that generates optimal and reliable results with even highly processed, fatty, or acidic foods, or foods with high or low DNA contents or high inhibitor contents (see the figures "High yields of DNA with minimal inhibitor carryover").

mericon DNA Bacteria and DNA Bacteria Plus Kits

The thermal lysis protocol of the mericon DNA Bacteria Kit and the mechanical lysis protocol of the mericon DNA Bacteria Plus Kit are similarly efficient for the extraction of DNA from Gram-negative bacteria, while the mechanical lysis protocol is more effective for the extraction of DNA from Gram-positive bacteria (see the figure "Efficient bacterial DNA extraction").

Fundamento
DNeasy mericon Food Kit

This kit uses an improved cetyltrimethylammonium bromide (CTAB) extraction of total cellular nucleic acids. The nonionic detergent CTAB is widely used for efficient extraction of total cellular nucleic acids from a wide range of tissue types. Depending on the salt conditions, CTAB may complex with cellular nucleic acids (low-salt conditions) or complex with cellular inhibitors, such as polysaccharides, proteins, and plant metabolites (high-salt conditions; as found in the Food Lysis Buffer).

With fewer workflow steps than conventional CTAB protocols, up to 30 samples can be processed in 2.5 hours with the DNeasy mericon Food Kit (see the figure "Efficient and rapid DNA extraction").

The optimized protocol uses CTAB in combination with Proteinase K to first digest compact tissue and then to subsequently precipitate proteins with simultaneous precipitation of other cellular and food-derived inhibitors.

Inhibitors are precipitated by centrifugation, while the extracted DNA remains in solution. In the subsequent chloroform extraction, any remaining CTAB-protein, CTAB-debris, or CTAB-polysaccharide complex not precipitated is removed together with other lipophilic inhibitors by extraction into the organic chloroform phase. Only the aqueous phase containing the DNA and significantly depleted inhibitors is processed further. This phase is mixed with binding buffer (to adjust binding conditions) and applied to the QIAquick Spin Columns. The resulting DNA is immediately ready for use in a downstream mericon real-time PCR assay.

mericon DNA Bacteria Kit and mericon DNA Bacteria Plus Kit

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.

The thermal lysis protocol of 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.). The mechanical lysis protocol of the mericon DNA Bacteria Plus Kit creates the appropriate conditions for bacterial extraction from Gram-positive bacteria, such as Listeria spp.

The 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.

Procedimento
DNeasy mericon Food Kit

DNA is extracted from a homogenized food sample using a straightforward protocol that removes inhibitors and yields pure DNA that is suitable for downstream applications, such as real-time PCR ingredient authentication or GMO detection.

mericon DNA Bacteria Kit and mericon DNA Bacteria Plus Kit

Bacterial DNA is extracted from a bacterial enrichment culture using a protocol based on mechanical lysis (using beads) or thermal lysis (by boiling). Both protocols deliver a bacterial DNA suspension suitable for real-time PCR pathogen detection.

Aplicações

The DNeasy mericon Food Kit is for isolation of DNA from raw or processed foods for downstream applications, such as ingredient authentication.

mericon Bacteria and Bacteria Plus Kits are for the isolation of bacterial DNA from enrichment cultures of food and animal feed.

Característica
Especificações
Applications PCR, real-time, genotyping
Elution volume 100 µl (mericon DNA Bacteria and DNA Bacteria Plus Kit), 150 µl (DNeasy mericon Food Kit)
Format spin columns (DNeasy mericon food Kit), microcentrifuge tube (DNA Bacteria Kit), icrocentrifuge tube with glass beads (DNA Bacteria Plus Kit)
Main sample type fod, e.g. chocolate, ketchup, cereals (DNeasy mericon Food Kit), bacterial culture from food (DNA Bacteria Kit and DNA Bacteria Plus Kt)
Processing manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein DNA
Sample amount 1 ml culture (DNA Bacteria and Bacteria Plus Kit), 2 g (DNeasy mericon Food Kit)
Technology silica technology (DNeasy mericon food Kit), boiling lysis (DNA Bacteria Kit), disruption with glass beads (DNA Bacteria Plus Kit)
Time per run or per prep 20 minutes - 1 hour (DNeasy mericon Food Kit)
Yield depends on DNA contents of the sample

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