mericon Sample Preparation and Bacteria Kits

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

Features

  • Extraction of high-quality nucleic acids from a range of starting materials
  • Minimal carryover of PCR inhibitors
  • Isolated, pure DNA suitable for real-time PCR applications 
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mericon DNA Bacteria Kit (100)

Cat. No. / ID: 69525

Fast Lysis Buffer
Kit
mericon DNA Bacteria Kit
mericon DNA Bacteria (Plus) Kit
DNeasy mericon Food Kit
mericon Sample Preparation and Bacteria Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Product Details

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.

Performance

DNeasy mericon Food Kit

During production, food is subjected to heat, irradiation, high pressure, and pH changes, leading 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 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").

See figures

Principle

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 the 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 digest compact tissue and 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 mericon DNA Bacteria Plus Kit maximizes lysis efficiency, stabilizes the extracted DNA and removes inhibitors that may interfere with downstream applications.

See figures

Procedure

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.

Applications

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 bacterial DNA isolation from enrichment cultures of food and animal feed.

Features Specifications
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

Supporting data and figures

Resources

Safety Data Sheets (2)
Download Safety Data Sheets for QIAGEN product components.
Download Safety Data Sheets for QIAGEN product components.

FAQ

Are there important considerations for plasma generation and urine handling?

It is strongly advised to follow the recommendations for preparing sample material provided in the corresponding Protocol Sheet to ensure reliable results.

Plasma: It is recommended to perform plasma separation immediately after blood collection when using EDTA or citrate as anticoagulant to prevent the release of genomic DNA into the plasma fraction.

Urine: Because circulating cell-free DNA in non-stabilized urine samples is rapidly degraded after sample collection due to high nuclease activity, eluates may contain no DNA or exhibit low DNA concentration. Therefore, it is recommended to stabilize urine samples. Even when using stabilized urine, it is recommended to perform a centrifugation step immediately after stabilization to prevent the release of genomic DNA from cells. Alternatively, non-stabilized urine samples can be processed immediately after collection and centrifugation using ATL-pretreatment and automated DNA extraction as described in the corresponding Protocol Sheet.

FAQ ID - 3699
When would I use the mericon DNA Bacteria Kit vs the mericon DNA Bacteria Plus Kit?
The mericon DNA Bacteria Kit is intended for the extraction of total genomic DNA from Gram-negative bacteria from pathogen enrichment cultures. The mericon DNA Bacteria Plus Kit is intended for the extraction of total genomic DNA from Gram-positive bacteria from pathogen enrichment cultures, or other pathogens that are difficult to lyse.
FAQ ID - 3348
Why is my 260/280 ratio low after using the DNeasy mericon Food Kit?
With complex food samples, a classical evaluation of DNA purity and protein content by A260/A280 ratio is not a significant criterion. Due to inevitable traces of food matrix in the sample background, a spectroscopic evaluation of DNA or protein absorptions is highly unreliable, and we have found that there is not a correlation between 260/280 and 260/230 ratios and PCR performance. Even DNA with Low 260/280 and 260/230 can produce good PCR results.

FAQ ID - 3349
Does salt concentration in the food matter when using the DNeasy mericon Food Kit?
We have not observed any effect of NaCl concentration in the food on the effectiveness of the DNeasy mericon Food Kit.
FAQ ID - 3346
CAn I use milk or other liquids with the DNeasy mericon Food kit? What volume would I use?
Liquids can be used. Use 2 ml (for 2 g protocol) or 200 ul (for 200 mg protocol) and mix it with the Food Lysis Buffer and Proteinase K solution as in the normal protocol.
FAQ ID - 3350
Why does the DNeasy mericon Food Kit use a QIAquick column and Buffer PB rather than a DNeasy column and Buffer AL, which might be expected since the kit isolates genomic DNA?
During kit development, we determined that with the chemistry used in the kit and the tested sample material, the QIAquick column together with the buffer PB had the best performance.
FAQ ID - 3347