BIOTECON Diagnostics DNA / RNA Extraction DNA / RNA Extraction – Key to Successful Analysis
StarPrep Kits ShortPrep Kits Sample Preparation Kits Magnetic Preparation Kits
Fast and easy lysis by mixing samples with buffer High purity DNA/RNA clean-up, manual column based or automated magnetic bead based
Reliable analysis of food and feed samples with real-time extraction kits have been optimized in combination with PCR require appropriate sample preparation. Optimized our foodproof® real-time PCR Detection, Identification and and validated methods for the extraction of nucleic acids Quantification Kits. The isolated DNA/RNA can be used from any kind of biological material are of high importance. directly in all our real-time PCR assays. The processed sample must be free of contaminants, especially inhibitors, while retaining small amounts of Direct Lysis, Column Based or Automated the target’s genetic information. The extraction method The foodproof® ShortPrep, StarPrep, Sample Preparation should be as fast and convenient as possible without and Magnetic Preparation Kits assist you in efficiently compromising safety or reproducibility. working with all types of sample matrices, including difficult complex samples like e.g. cheeses, spices or cacao. Optimized Combination of Extraction Kit and PCR Assay BIOTECON Diagnostics has developed a broad range of With fast, straight-forward and easy-to-use solutions, you fast, safe and easy-to-use DNA and RNA extraction kits for can overcome the challenges of complex matrices, PCR all kinds of food, beverage or environmental samples. Our inhibitors and hidden targets.
2 DNA / RNA Extraction – Key to Successful Analysis
Table 1: Comparison of foodproof® extraction kits for food and beverage-relevant parameters
Rapid Highly purified Target Bead based/ Bulk Prefilled Column based Automated
Sample Magnetic Gram-negative bacteria StarPrep One* ShortPrep I Preparation I Preparation I, V
Sample Magnetic Gram-positive bacteria StarPrep Two* ShortPrep II Preparation II Preparation II
Enterobacteriaceae, Magnetic StarPrep One - - Cronobacter Preparation IV
Yeast & Mold, Dekkera, S. StarPrep Two - - - cerevisiae var. diastaticus
Vibrio, STEC, StarPrep Three* - - - Beer spoilage bacteria
Sample Magnetic GMOs, Animal ID, Allergens StarPrep Five** - Preparation III Preparation III
Sample Magnetic Viruses - - Preparation IV Preparation Kit VI
* High-throughput protocol available. See 8-strip DNA extraction protocols. **Only for Animal ID
3 StarPrep Kits: Versatile
WORKFLOW e.g. StarPrep One Enjoy convenient and flexible DNA StarPrep can be used for rapid DNA extraction: the foodproof® StarPrep extraction from enrichment cultures, Kits are designed for quick and direct and rinse samples. The reagents ADD SAMPLE easy extraction of DNA from food contain no hazardous chemicals or CENTRIFUGE 5 min DISCARD SUPERNATANT and beverage samples by heat- compounds, unlike other sample induced lysis. StarPrep is a cost- preparation methods. The entire DNA effective, versatile solution for nearly preparation can be performed in a ADD LYSIS BUFFER all kind of foods, with optimized lysis single tube, minimizing handling steps, reagents and protocols for different reducing risk of cross-contamination applications. and exposure to biohazardous material. StarPrep One Kit Bulk solution for fast and flexible DNA extraction MIX Table 2: Overview of foodproof® StarPrep Kits
foodproof® StarPrep One foodproof® StarPrep Two foodproof® StarPrep Three foodproof® StarPrep Five
Gram-positive and other Vibrio, INCUBATE bacteria, e.g. Listeria, 10 min STEC, beer spoilage Gram-negative bacteria, Campylobacter, E. coli Targets bacteria, Salmonella Animal ID e.g. Salmonella O157, Alicyclobacillus; spp. plus Enteritidis & Yeast and Mold, Typhimurium Legionella MIX Suitable for a wide range of food, animal feed and Enrichment cultures, Matrices Meat, processed food environmental samples beverage samples Mechanical disruption and Principle Heat-induced lysis Heat-induced lysis Heat-induced lysis CENTRIFUGE heat-induced lysis 1 min Isolations 100 / 500 96 96 96
Additional - Resuspension Reagent - - SUPERNATANT Products FOR DETECTION MicroVal, AOAC, Approvals AOAC, NordVal and DAFF AOAC-RI - NordVal and DAFF
4 ShortPrep Kits: Ready-to-Use
WORKFLOW The fast and easy DNA extraction method for your enriched e.g. ShortPrep I samples: the foodproof® ShortPrep Kits are designed for quick and easy DNA extraction of bacterial DNA to be used directly in PCR. Simply add enriched sample to pre-aliquoted buffer, and in less than 30 minutes, samples from bacterial enrichment cultures are ready for PCR. The entire extraction ADD SAMPLE is performed in a single tube, minimizing handling steps and to prefilled lysis buffer reducing risk of cross-contamination. The lysis buffers are free of dangerous reagents. The kits are validated and certified by internationally MIX recognized institutions such as AOAC, NordVal and DAFF, and can be used for a variety of foods, like raw materials or processed food, environmental samples and pet foods. ShortPrep I Kit Prefilled tubes for fast DNA extraction INCUBATE in less than 30 minutes 10 min Table 3: Overview of foodproof® ShortPrep Kits
foodproof® ShortPrep I foodproof® ShortPrep II MIX Gram-positive and other bacteria, e.g. Listeria, Targets Gram-negative bacteria, e.g. Salmonella Campylobacter and Alicyclobacillus
Matrices* Suitable for a wide range of food, animal feed and environmental samples CENTRIFUGE Principle Heat-induced lysis Mechanical disruption and heat-induced lysis 1 min
Convenience 96 individual ready-to-use reaction mixes for maximum efficiency and easy DNA extraction
Approvals AOAC, NordVal and DAFF SUPERNATANT FOR DETECTION
* DNA extracts from some raw materials of plant origin (e.g. spices) and from some beverage samples may still contain substances that interfere with PCR amplification or with fluorescent signal detection. For such materials, preparation and purification with the BIOTECON Diagnostics’ foodproof Sample Preparation Kit I and II are recommended.
5 Sample Preparation Kits: High Quality Purification
WORKFLOW e.g. Sample Preparation III Kit* Get rid of unwanted PCR contaminants and inhibitors: the foodproof® Sample Preparation Kits are designed to isolate and purify DNA or RNA even from challenging ADD EXTRACTION BUFFER matrices containing high amounts of PCR inhibitors. Kit to sample reagents and consumables are optimized to obtain the best results for each parameter. Protocols are easy to Sample Preparation III Kit MIX follow and implement in daily routine use. The quality of the INCUBATE 30 min Column-based DNA obtained with the kits is suitable for any application extraction for CENTRIFUGE 10 min using real-time PCR. highly purified DNA
TRANSFER SUPERNATANT
ADD BINDING BUFFER & PROTEINASE K Table 4: Overview of foodproof® Sample Preparation Kits INCUBATE 10 min ADD ISOPROPANOL foodproof® Sample foodproof® Sample foodproof® Sample foodproof® Sample Preparation I Kit Preparation II Kit Preparation III Kit Preparation IV Kit TRANSFER SUPERNATANT to filter column Gram-positive and other Genetically modified CENTRIFUGE 1 min Gram-negative bacteria, bacteria, e.g. Listeria, organisms (GMOs), Viruses Targets DISCARD FLOW THROUGH e.g. Salmonella Campylobacter and animal identification (e.g. Norovirus) REPEAT ONCE Alicyclobacillus allergens
ADD WASH BUFFER Matrices Suitable for a wide range of food, beverages, animal feed and environmental samples CENTRIFUGE 1 min The RNA is bound to a DISCARD FLOW THROUGH The DNA is bound to a glass fiber filter, washed thoroughly and the pure DNA is eluted Principle glass fiber filter, washed REPEAT ONCE from the columns and eluted
ADD ELUTION BUFFER Isolation of highly purified DNA, virtually free of other cellular components and PCR inhibitors CENTRIFUGE 1 min Isolation of viral RNA, Features Suitable for matrices containing high amount of PCR inhibitors FLOW THROUGH process control included FOR DETECTION Sample Preparation Kit II and Sample Preparation Kit III include a lysozym and Proteinase K digestion step, respectively, for maximum performance * Workflow for GMOs & allergens; animal ID slightly different protocol
6 Sample Preparation Kits: High Quality Purification Magnetic Preparation Kits: Automated Purification
foodproof RoboPrep® BIOTECON Diagnostics offers a variety of solutions The foodproof RoboPrep® 32 is perfect for automated high-throughput molecular testing for for labs with low to medium sample pathogens, GMOs, animal identification and allergens. throughput allowing the automated The foodproof® Magnetic Preparation Kits are based on extraction of up to 32 samples magnetic bead technology, providing ultra-pure, ready- simultaneously. Despite being small and to-use genomic DNA of bacterial, plant or animal origin economic, the RoboPrep® 32 offers the for enhanced performance in sensitive real-time PCR Magnetic Preparation Kit I full spectrum in terms of efficiency and applications. The process proceeds with DNA being bound Bead-based convenience. Nucleic acid extraction with to magnetic beads and washed thoroughly through multiple DNA extraction magnetic beads is combined with heating steps. In the final step, highly purified DNA is eluted from for automation blocks enhancing sample lysis and the beads and is ready for use. elution. The result is highly purified DNA / Table 5: Overview of foodproof® Magnetic Preparation Kits RNA, free of inhibitory components, which can be directly used for PCR assays. MPK I MPK II MPK III MPK IV MPK V MPK VI
® Order no. S 400 11 L S 400 12 L S 400 13.2 L S 400 15 L S 400 19 L S 400 20.1 L The foodproofRoboPrep 32 is optimized for foodproof® Magnetic Preparation Kits, Entero- Gram-negative Gram-positive bacteriaceae, however, any other kind of protocol can GMOs, animal ID, Targets bacteria, e.g. bacteria, Cronobacter Salmonella spp. Viruses allergens be easily implemented via the intuitive Salmonella spp. e.g. Listeria spp., Salmonella spp., touch display of the device. Serum, plasma, Matrices Food and feed Food and feed Food and feed Infant formula Food feces and swab samples AOAC and Approvals - - MicroVal - - MicroVal
RoboPrep Fusion®, RoboPrep Fusion® RoboPrep Fusion®, RoboPrep Fusion®, RoboPrep Fusion®, KingFisherTM Flex, KingFisherTM Flex, TM KingFisherTM Flex KingFisherTM Flex, Instruments KingFisher Flex RoboPrep® 32, RoboPrep® 32, RoboPrep® 32, RoboPrep® 96 RoboPrep® 96 RoboPrep® 96
7 Reagent D: Elimination of DNA from dead cells
PRINCIPLE Matrices like powdered infant formula or Elimination Procedure 1. Reagent D Treatment processed seafood may contain a high Reagent D contains light-sensitive background of dead (inactive) target substances that selectively penetrate cells, which may cause false-positive cell membranes of dead cells. Exposure results or distort quantitative PCR to visible light leads to covalent binding
live cells dead cells assays. Reagent D is highly selective of Reagent D to DNA and prevents the in penetrating only membranes of masked DNA from being amplified in dead cells and can be used for live/ the PCR. Reagent D has been validated dead differentiation to avoid false- in combination with the foodproof® positive results. StarPrep Kits. Reagent D Kit 2. After Extraction Processing takes only a few minutes For elimination of false-positive results prior to performing the DNA extraction. from dead cells to avoid false-positive results
foodproof® D-Light light dose with high power LEDs and 3. Real-time PCR Run As a validated source of light for proper optical alignment guarantees DNA of live cells DNA of dead cells the photoactivation of Reagent D, reproducible results. The workflow is the stand-alone CE/FCC certified simple and safe: no cooling needed, as ® amplification amplification blocked foodproof D-Light is a compact no heat is generated. benchtop solution that fits in every lab. The combination of the foodproof® Up to 96 samples can be processed D-Light and Reagent D, foodproof® without Reagent D with Reagent D simultaneously. SBS format allows DNA extraction kits and subsequent
Intensity Intensity different adapters and microplates to real-time PCR kits and all assay- live cells be used. A pre-programmed incubation required laboratory equipment live + dead dead cells cells starts automatically after insertion of represents a true one-stop shop Cycles Cycles the samples. A constant and uniform solution for the customer.
8 Reagent D: Elimination of DNA from dead cells 8-Strip DNA Extraction Protocols
WORKFLOW ® For efficient manual handling of large sample numbers, foodproof StarPrep Two 8-Strip Kit e.g. StarPrep One 8-Strip BIOTECON Diagnostics has developed time-saving high- In less than 30 minutes, the DNA of gram-positive bacteria throughput protocols for DNA extraction for many applications, like Listeria monocytogenes can be prepared from up to 800 including Salmonella and Listeria enrichment cultures and µl food enrichments for direct use in any PCR application. The ADD SAMPLE yeast and mold samples. protocols include a disruption step. CENTRIFUGE 10 min The usage of an 8-channel pipette reduces the number DISCARD SUPERNATANT ® of handling steps, saving time and effort. The entire DNA foodproof StarPrep Six 8-Strip Kit (for Yeast and Mold) preparation is done in a single tube: Transfer steps of samples This is a quick and easy high-throughput DNA extraction method for yeast and mold detection in foods. The protocol ADD DILUTED are not necessary, thus cross-contamination risks are LYSIS BUFFER includes a differentiation of live and dead cells using Reagent minimized. The cells are lysed by heat and, depending on target MIX organism, by mechanical disruption. After centrifugation, the D and is validated with a broad range of dairy products. to resuspend pellet DNA is in the supernatant and ready to be used in any PCR Table 6: Overview of additional equipment application. StarPrep StarPrep StarPrep StarPrep One Two Three Six foodproof® StarPrep One and StarPrep Three 8-Strip Kits Instrument 8-Strip 8-Strip 8-Strip 8-Strip INCUBATE ® 10 min The foodproof StarPrep One and StarPrep Three 8-Strip Kit Kit Kit Kit Kits are designed for a rapid and easy extraction of DNA from Magnetic X - X - gram-negative bacteria like Salmonella or Cronobacter in food Stirrer enrichments. The straightforward, easy-to-follow protocol Heating Block X X X X saves time while maintaining high sensitivity. Centrifuge X X X X CENTRIFUGE 5 min Mechanical Cell - X - X Disruption Multichannel X X X X Pipette SUPERNATANT FOR DETECTION Consumables X included X included
9 Basic Lab Instruments
BIOTECON Diagnostics provides its customers a “one- nucleic acid extraction and make full use of our real-time stop shop” solution. We proudly offer instruments and PCR kits. This select equipment has been thoroughly laboratory equipment necessary to perform our high quality tested by our team to guarantee quality performance.
Disruptor Genie® Micro Star 17 microcentrifuge Heating-Thermomixer MHR 13 Ideal for cell disruption/homo- Powerful (17,000 ×g), safe, compact, All-in-one: heating, cooling, mixing, genization of bacteria and pellet versatile, convenient and ventilated timer with one single unit resuspensions centrifuge 30 different exchange blocks Hands-free automatically timed Easy operation, intuitive controls, available (e.g. for the use of micro disruption of up to 12 microtubes bright and easy to read displays test tubes, test tubes, centrifugal Patented multi-directional action Designed to accelerate the routine tubes, cuvettes or PCR plates, agitates and vortexes at high-speed sample preparation process microtiter or deep-well plates) producing greater yields in less time Special ‘click’ biocontainment rotor lid State-of-the-art touch panel for Suitable for use in cold rooms or in keeps the user and the samples safe better handling and cleaning incubators Supplied with 24-place rotor for Possible to program up to 9 Can be also used as a vortexer 1.5/2.0 ml tubes and aerosol-tight complete programs with up to 30 rotor lid steps
10 Basic Lab Instruments
Table 7: Basic laboratory equipment required for performing nucleic acid extraction Magnetic Heating Reagent D & RNA/DNA Extraction Kit Disruptor Vortex Centrifuge Stirrer Block D-Light
foodproof® StarPrep One Kit X 1.5 ml Block - EbC* X ≥13,000 x g
Yeast & Mold, foodproof® StarPrep Two Kit X 1.5/2 ml Block X X ≥13,000 x g Legionella
foodproof® StarPrep Three Kit X 1.5 ml Block - Vibrio X ≥13,000 x g
foodproof® StarPrep Five Kit - 2 ml Block - - - ≥13.000 x g
foodproof® ShortPrep I Kit - 1.5 ml Block - - X ≥8,000 x g
foodproof® ShortPrep II Kit - 1.5 ml Block X - X ≥13,000 x g
foodproof® ShortPrep III Kit - 1.5 ml Block - - X ≥15,000 x g
foodproof® Sample Preparation Kit I - 1.5 ml Block - - X ≥13,000 x g
foodproof® Sample Preparation Kit II - 1.5 ml Block - - X ≥13,000 x g
foodproof® Sample Preparation Kit III - 2 ml Block - - X ≥13,000 x g
foodproof® Sample Preparation Kit IV - - - - X ≥20,000 x g
foodproof® Magnetic Preparation Kit I ------
foodproof® Magnetic Preparation Kit II - - - - - ≥2,250 x g**
2 ml Block, foodproof® Magnetic Preparation Kit III - - - X - mix
foodproof® Magnetic Preparation Kit IV - - - EbC* - -
foodproof® Magnetic Preparation Kit V ------
foodproof® Magnetic Preparation Kit VI ------
* Enterbacteriaceae plus Cronobacter ** Centrifuge for deep-well plates
11 www.bc-diagnostics.com [email protected] Fax: (0)331-2300-200 Phone: +49 Germany 14473 Potsdam Hermannswerder 17 BIOTECON DiagnosticsGmbH www.product-finder.bc-diagnostics.com Find theproduct youare lookingforat: INSTRUMENTS For publications,references and certificates, pleasevisitourwebsite: +49 (0)331-2300-299 www.bc-diagnostics.com EXTRACTION PCR
BR-DNAEX-EN V02/2021 Product List S 40022 S 40020.1L S 40019L S 40018L S 40015L S 40013.2 L L S 40012 S 40011L S 40011 S 40021 S 40018 S 40017L S 40008.1 S 40014L S 40007L S 40007 S 40016 S 40006.1 S 40005 S 40004 S 40003 S 40002 S 40001 DNA ExtractionKits
food food food food food food food food food food food food food food food food food food food food food food food proof proof proof proof proof proof proof proof proof proof proof proof proof proof proof proof proof proof proof proof proof proof proof ® ® ® ® ® ® ® ® ® ® ® ® ® ® ® ® ® ® ® ® ® ® ® StarPrep Six8-StripKit,192reactions MagneticPreparation KitVI,480reactions MagneticPreparation KitV, 480reactions StarPrep Three 8-StripKit,480reactions MagneticPreparation KitIV, 480reactions MagneticPreparation KitIII,480reactions MagneticPreparation KitII,480reactions MagneticPreparation KitI,480reactions MagneticPreparation KitI,96reactions StarPrep Five,96reactions StarPrep Three Kit,96reactions StarPrep Two 8-StripKit,480reactions StarPrep Two Kit,96reactions StarPrep One8-StripKit,480reactions StarPrep OneKit,500reactions StarPrep OneKit,100reactions SamplePreparation KitIV, 50reactions SamplePreparation KitIII,50reactions SamplePreparation KitII,100reactions SamplePreparation KitI,100reactions ShortPrep IIIKit,96reactions ShortPrep IIKit,96reactions ShortPrep IKit,96reactions
D 22032 06 600 Z 01 220 D Automation A widerrangeavailableonrequest D 11027 39 110 D D 11038 D 11044 D 11010 D 10003 Basic LabInstruments A 50018 A 50014 A 50012 A 50002S A 50002 Additionals
food KingFisher KingFisher Magnetic Stirrer deep-well plates TH 21Exchange heating block for TH 21HeatingBlock Heating-Thermomixer MHR13 Micro Star17microcentrifuge Disruptor Genie Rinse Buffer, 110ml Resuspension Reagent,35ml Reagent P, 550reactions Reagent D,15ml Reagent D,30ml proof RoboPrep TM TM FlexConsumablePack Flex ® ® 32