QIAGEN Plasmid Plus Spin Columns QIAfilter Midi Cartridges Caps for QIAfilter Tube Extenders (20 ml) Collection Tubes (2 ml) Buffer P1 Buffer P2 Buffer S3 Buffer ETR Buffer BB Buffer PE (concentrate) Buffer EB RNase A* LyseBlue Handbook 12943 25 25 Midi columns 25 25 25 25 1 x 110 m. Can the QIAcube be connected to a laboratory information management system? What is the expected level of endotoxins in plasmid DNA purified with QIAprep spin miniprep kit? Easy automation. 0000023455 00000 n
New software or software updates can beaccessed via the QIAcube Web Portal at www.qiagen.com/MyQIAcube. No change of chemistry is needed, so startup is fast, results are immediate, and performance is comparable to the manual procedure. What are the additional plasmid bands I see on my gel? However, carbohydrate contamination may also be observed when using other strains. These come with spin columns and tubes already loaded onto the rotor, so you don't have to . Plasmid pCMV DNA was prepared using the indicated preparation method with standard and high-yield (HY) protocol for QIAGEN Plasmid. Includes 1 Preventive Maintenance or Inspection Service during the Premium Agreement period. AirPore Tape Sheets. However, this buffer can be purchased separately: Do you have a protocol for obtaining > 20 g plasmid DNA using the QIAprep Spin Miniprep Kit? 0000012460 00000 n
Therefore, the wells of the DNeasy 96 plate are at great risk of clogging. You can stop a QIAcube protocol if there is an emergency by pressing Cancel. %PDF-1.5
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High yields, fast procedures, as well as convenient and flexible processing options are just some of the benefits experienced with this unique technology. We do not have this part available for replacement. Can the QIAvac 24 Plus manifold be autoclaved? QIAsphere gives full connectivity so you can step out of the lab to ponder on your research while keeping an eye on your instrument run status. Filter-Tips, 1000 L (1024), cat no. QIAGEN resin is a macroporous silica-based resin with a high density of diethylaminoethyl (DEAE) groups that was developed exclusively for isolation of nucleic acids. Report files can be automatically transferred from your QIAcube Connect using the QIAsphere Export Tool. This handling error leads to inefficient cell lysis, and incomplete precipitation of SDS, cell debris, and genomic DNA. When using the silica-based QIAprep Spin Miniprep Kit, a protocol is contained in the QIAprep Miniprep Handbook, in Appendix C: Special Applications. The last protocol step performed by the QIAcubewill bedisplayed in the touch screen. For manual extraction, supplementary protocols are available for the QIAamp DNA Mini and Gentra Puregene Cell Kits (see resources section). Viscosity of the lysates varies from sample to sample. The resulting highly concentrated DNA is ready for immediate use in subsequent applications. The last protocol step performed by the QIAcube will be displayed on the touchscreen. This product is not intended for the diagnosis, prevention, or treatment of a disease. Average endotoxin levels that we have observed for Silica gel slurry is around 1200 EU/g DNA. The QIAGEN Plasmid Plus Kits provide transfection-grade plasmid DNA highly suited for all applications, such as: We do not recommend using a vacuum manifold for DNA extraction using the QIAamp 96 DNA Blood Kit because: Please contact Nalgene for ordering. (Endotoxin levels can vary depending on the host strain, culture media, and cell density. applications Details on buffer preparation and storage are presented in Appendix B of the QIAGEN Plasmid Purification Handbook. Do not unzip the files. See: http://www.qiagen.com/products/catalogpagecontrols/header/qiacube/360/default.aspxfor a demonstration on how the QIAcube is used with the RNeasy Mini Kit for automated purification of RNA. 0000025472 00000 n
LyseBlue reagent is provided inthe following QIAGEN plasmid kits: Find out which origin of replication your plasmid contains, andlook at the table below for classification into high-copy or low-copy types. Preparation of LB medium: Dissolve 10 g tryptone, 5 g yeast extract, and 10 g NaCl in 800 ml dH2O. Instrument, 1-year warranty on Increase the efficiency and productivity of your lab. Filter the solution through a 0.2 m sterile filter using a syringe to give a final volume of 6 ml Buffer MP. 0000007226 00000 n
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With LyseBlue reagent for lysis control, can I now process more bacterial culture and overload the columns? Endotoxins, also known as lipopolysaccharides or LPS, are cell-membrane components of Gram-negative bacteria such as E. coli and are released during the lysis step of plasmid purification. plasmid DNAfor Download more information. What applications are offered on the QIAcube? The most common cause of this problem isover-growth of bacterial cultures. We would expectthe enzymeto have some residual activity. The QIAGEN Plasmid Plus chemistry is not compatible with Buffer ER from the Endofree Plasmid kits. Fast, inexpensive Note: The apparent power can exceed 650 VA for up to 2 seconds during the centrifuge acceleration and can reach an approximate value of 1000 VA.; The QIAcubeis equipped with an optical sensor, which determines the tip size and the number of tips loaded onto the worktable prior to starting a run. QIAGEN Plasmid Plus Kit Midi (25) Catalog. use in most downstream Storage temperature range: 20C to 60C (4F to 140F); Response time of next business day. QIAGEN has developed a wide range of silica gel membrane products that selectively bind either RNA or DNA and separate nucleic acids within certain size parameters. Useful hints and information on optimizing plasmid preparations can be found at the QIAGEN Plasmid Resource Center. 990352. For isolation of genomic, mitochondrial, bacterial, parasite or viral DNA, For purification of up to 20 g molecular biology grade plasmid DNA, For a more eco-friendly alternative to our standard kit for extracting up to 20 g molecular biology grade plasmid DNA, For automated purification of DNA from forensic and HID samples on the EZ1 Adv XL or EZ2 Connect Fx Instruments, For automated isolation of cell-free (cfDNA) DNA from human plasma or serum on EZ1 Advanced XL or EZ2 Connect, For isolation of free-circulating DNA and RNA from human plasma or serum, For the fastest, most convenient purification of up to 10 mg transfection-grade plasmid DNA, For purification of genomic, mitochondrial or viral DNA from blood and other body fluids. Fluorescent and radioactive sequencing (including capillary sequencing), ligation, cloning, transformation, etc. Buffer P1 with RNase A used in QIAGEN Plasmid Purification Kits should be fineat room temperature for a few days. Note: Do not rename or modify the protocol files. Prep 96 protocol'. LyseBlue reagentnow allows the user to monitor potential problems (insufficient bacterial cell resuspension and lysis as a consequence of overloading) early in the plasmid preparation process. How can I purchase replacement bottles, caps and tubing for my QIAvac Connecting System? When resuspending the cell pellet, vortexing longer or resuspending the pellet by pipetting upand down can help. Do you want to expand your application range or further streamline workflows? A convenient tool to build experimental workflows and find products to match your needs. Note: If a protocol run is stopped, the run cannot be restarted. QIAamp PowerFecal Pro DNA QIAcube Kit (240), RNA purification: A direct comparison of QIAcube Connect with the Promega Maxwell RSC, Extraction of microbiome DNA using the DNeasy PowerSoil Pro Kit manually and automated on the QIAcube Connect and QIAcube, Decontamination procedure using the UV LED source from QIAcube Connect, Improved differential cell lysis and extraction using an automated protocol on the QIAcube Instrument, QIAcube Connect Safety Instructions and Quick Start Guide, Request QIAcube Connect Customized Protocol, Ensure success with QIAGEN services for QIAcube Connect, Ensure success with QIAGEN Service Solutions for QIAcube Connect, Redefine the boundaries of automated sample processing, Automated differential wash protocol using the QIAcube Connect and the EZ1 DNA Investigator Kit, QIAcube Connect Instrument Software 1.1.1, QIAcube Connect System Technical Information, http://www.qiagen.com/products/catalogpagecontrols/header/qiacube/360/default.aspx, Power: 100240 V AC, 50/60 Hz, 650 VA Mains supply voltage fluctuations are not to exceed 10% of nominal supply voltages. ); All standard protocols in the expanding range can also be downloaded free of charge. Plasmid DNA prepared with QIAGEN Plasmid Plus 96 Miniprep and BioRobot Kits resulted in highly efficient transfection into sensitive cell lines. Yes,it is possible to isolateplasmid DNAfrom mammalian cells using the QIAprep Spin Miniprep kit . The TRACKMAN Connected system guides researchers through the QIAprep Spin Miniprep protocols while automatically adjusting the Bluetooth-enabled PIPETMAN M Connected pipette settings. Explore high-quality enzymes; now available as individual products. When purifying DNA, it is critical to use an optimized method for your sample type. ), height 86 cm (34 in. Note: If a protocol run is stopped, the run cannot be restarted. Clumps that occur after addition of Buffer P2in a bacterial lysatecontaining LyseBlue reagent indicatepoor resuspension of the bacterial cell pellet in Buffer P1. For spin-column or 96-well extraction of total DNA from animal blood and tissues and from cells, yeast, bacteria, or viruses. You can also access this informationon our Plasmid Resource Pages. Nucleic acids are isolated from lysates through binding to the magnetic particles in the presence of a chaotropic salt, which removes water from hydrated molecules in solution. Can I program my own protocols for the QIAcube? They include a silica resin that selectively binds DNA or RNA relying on the factors involved in the extraction method. Through a simple color change, this unique buffer additive ensures that optimum cell lysis and neutralization is achieved, maximizing yields. What should I do about that? Therefore, the wells of the QIAamp 96 plate are at great risk of clogging. 0000008930 00000 n
The final protocol for large scale Mirapreps was tested with GeneJet, QIAGEN, and Sigma GenElute spin column Miniprep kits, using the resuspension, lysis, neutralization and wash buffers provided (a step-by step protocol is provided in S1 File). The article in QIAGEN News 1995 No. What are spin columns? Assessing unmodified 70-mer oligonucleotide probe performance on glass-slide microarrays. ), depth 62 cm (24.4 in. How can I load new protocols onto the QIAcube? Is it possible to elute plasmid DNA from the QIAprep Spin Miniprep columns with buffer containing Potassium Phosphate? Molecular assays require high-quality nucleic acids to obtain reliable results. QIAcube Connect is QIAsphere-ready. RNA to the RNeasy spin column membrane, resulting in lower RNA yield and purity. ); Using our spin columns as an OEM component will help you ease the regulatory compliance burden while our global reach can simplify your logistics . GelPilot Loading Dye contains 3 tracking dyes (xylene cyanol, bromophenol blue, and orange G) to facilitate the optimization of agarose gel run time and prevent smaller DNA fragments migrating too far (see figure "GelPilot Loading Dye"). Press. How can I get software updates for the QIAcube? Part numbers from Nalgene: - 10 liter waste container (Nalgene Heavy Duty Vacuum Carboy [PP], cat. QIAGEN Plasmid Plus technology delivers the same performance and quality as anion-exchange technology. transfection grade DNA for These let the QIAcube Connect fully automate more than 80 QIAGEN spin column kits. One on-site Preventive Maintenance or Inspection Service visit for the QIAcube Connect, including travel, labor and parts. The purified DNA can also be eluted in TE (10 mM Tris-Cl, 1 mM EDTA, pH 8.0), but the EDTA may inhibit subsequent enzymatic reactions. QIAfilter cartridges provided in QIAGEN Plasmid Plus Kits enable rapid lysate clearing. Looking for a quick way to design experiments? Compatible OS: Windows 8, Windows 7, Windows Vista, Windows XP (SP3 or later); Mac OS x 10,1 or later; The high-copy plasmids listed here contain mutated versions of this origin. Do you have a replacement part available, so I can replace the gasket? QIAGEN offers a wide range of specialized nucleic acid purification products based on four highly developed purification technologies: solid-phase, anion-exchange technology; QIAGEN Plasmid Plus technology; silica gel membrane technology; and magnetic particle technology. Purified DNA can be used in restriction digestion (see figure "Complete digestion with various restriction enzymes"). During incubation, place QIAGEN Plasmid Plus spin columns into the vacuum pump. no. Operating humidity range: 10 to 90% (with no condensation); Quality Control As part of the stringent QIAGEN quality assurance program, the performance of QIAquick Spin Kits is monitored routinely and on a lot-to-lot basis. Dissolve 3.3 g citric acid monohydrate in 3 ml high-purity water at room temperature (21C) in a 10 ml tube. Since QIAGEN launched the worlds first kit for purification of plasmid DNA, technologies have been developed to make plasmid DNA preparation even more convenient and reduce endotoxin levels in line with scientists needs in the laboratory. Some bacterial strains, such as TG1 and JM100, naturally produce a high level of carbohydrates. Qiagen Plasmid Kit Plus Maxi Kit (12963, 12965) E.Z.N.A. Endo-Free Plasmid Maxi Kit (D6296) Isolate low endotoxin (<1EU/g) plasmid DNA from up to 200 mL bacterial cultures using spin maxi columns . Edited by: Fred M. Ausubel, Roger Brent, Robert E. Kingston, David D. Moore, J.G. HiSpeed Midi Tips, provided in the HiSpeed Plasmid Midi Kit, contain a newly developed anion-exchange resin. The quality of silica gel membranes used in QIAGEN products ensures consistent yields of high-purity nucleic acids. Explore the virtual tour to learn more and see this product in action. The samples must be processed manually. 0000025833 00000 n
For purification of up to 20 g molecular biology grade plasmid DNA, 24/7 automatic processing of online orders, Knowledgeable and professional Product & Technical Support. 0000002910 00000 n
The QIAcube can use partially used tip racks from previous runs, provided that the racks contain sufficient tips to process the number of samples loaded for the current run. Nucleic acids are adsorbed to the silica gel membrane in the presence of chaotropic salts, which remove water from hydrated molecules in solution. For automated purification of up to 20 g/well of molecular biology grade plasmid DNA. The total binding capacity of plasmid plus south column is 250 g of plasmid DNA. Where can I find a protocol for cleanup of already purified plasmid DNA? Can the QIAcube rotor and/or buckets be removed for cleaning? If the answer to any of these questions is yes, then QIAGENs trade-in/trade-up program is just right for you! no 990394, and disposable tips are required: To find out which filter-tips are required for a specific application, refer to the corresponding protocol sheet or the instructions on the QIAcube Connect touchscreen. Rotor-adapters that come with the kits are pre-loaded with spin columns and elution tubes, so they are ready to use right out of the box. A convenient tool to build experimental workflows and find products to match your needs. Slightly modified QIAGEN protocols or protocols fully customised to your specific requirements can be requested from QIAGEN. We are sorry but the page you are trying to reach is currently unavailable. High yields are ensured with QIAGEN Plasmid Plus Kits (QPP). Detect TB infection with confidence. No. We recommend that Buffer P1 with RNase A be stored in the refrigerator (28C). The plug-and-play QIAcube Connect simplifies purification by automating QIAGEN spin-column kits. For purification of low- copy plasmids and cosmids, large plasmids (>10 kb), and DNA prepared using other methods, refer to the recommendations on page 31. High-purity plasmid DNA eluted from QIAprep 2.0 Spin Columns is immediately ready to use there is no need to precipitate, concentrate, or desalt.To enable faster and more convenient sample processing and analysis, gel loading dye is provided in the kit. out of columns to catch the plasmid DNA and thus I have lots and lots of reagent over but have to buy an entire new kit if I want to continue to make. *Note: add Glucoseafter autoclaving the solution with the remaining ingredients, and letting it cool down. Viscosity of the lysates varies from sample to sample; therefore, the DNeasy mini spin columns are at great risk of clogging. Explore our digital magazine full of customer stories and videos on scientific breakthroughs and healthcare advances. Looking for a quick way to design experiments. 0000006665 00000 n
Versatile QIAprep 2.0 Spin Columns can be used either in microcentrifuges, on vacuum manifolds, or in the QIAcube (see figures . QIAcube Connect decontaminates your full worktable with built-in UV light (see "The QIAcube Connect UV light") to prevent carryover between samples (see "Genomic DNA (gDNA) barely detectable after UV light exposure"). Sometimes an additional band of denatured supercoiled DNA migrates just below the supercoiled form. The design and unique binding chemistry of the QIAGEN Plasmid Plus columns allow a simple bind-wash-elute procedure based on a novel chemistry. Environmental class: 1K2 (IEC 60721-3-1), Dimensions (hood closed): width 65 cm (25.6 in. Origins of replication and copy numbers of various plasmids and cosmids. Growth of bacterial cultures; Plasmid Copy Number. Try the Workflow Configurator. Explore targets and pathways in their scientific context, find and customize products to study them, analyze data and plan follow-up studies all in GeneGlobe. The nucleic acids are then efficiently washed and eluted under low- or no-salt conditions in small volumes of elution buffer. The plasmid DNA obtained is highly suitable for a multitude of applications, including transfection into sensitive cell lines. Please visit www.pall.com. After a wash step, pure nucleic acids are eluted under low- or no-salt conditions in small volumes, ready for immediate use without further concentration. Why do I get genomic DNA contamination in my plasmid prep? If you attempt to re-install an existing protocol, a message will appear indicating that not all protocols could be copied. Can I use your QIAvac 96 for extraction of DNA using the DNeasy 96 Blood & Tissue Kit? Do you need improved connectivity? No. However, please be aware that the plastic may change in color. How do I know if my plasmid is a high- or low copy number type? applications This will affect the purity and yield of samples. In partnership with My Green Lab, we've also assessed the environmental impact of the QIAprep Spin Miniprep Kit (250). However,below is a reference where cDNA was eluted from QIAquick PCR Purification Kit columns with potassium phosphate buffer (4 mM, pH 8.5), after replacing the wash buffer (PE) with 5 mMpotassium phosphate(pH 8.5) containing 80% ethanol: Wang HY, Malek RL, Kwitek AE, Greene AS, Luu TV, Behbahani B, Frank B, Quackenbush J, Lee NH. no.8000-0065). The QIAcube Connect System and 21 CFR Part 11 Regulations. For a detailed description on how to run and interpret an analytical gel, please see Appendix A in the QIAGEN Plasmid Purification Handbook: "Agarose Gel Analysis of the Purification Procedure", or visit the QIAGEN Plasmid Resource Center. To confirm that you want to stop the protocol run, press Yes. This precipitate will completely dissolve after addition of Buffer P2. For information on how to remove these parts, refer tothe QIAcube User Manual. Available in versatile . carry over, Leverage our purification and custom kitting know-how in your assay. formatsfor all scales of Optical power: 200-300mW, Handheld scanner; Includes a 10% discount on additional repair service during the Core Agreement period. Please be sure to shake Buffer P1 vigorously before use to completely resuspend LyseBlue particles. The procedure can be performed in 20 (Midi and Maxi), 40 (Mega), or 50 minutes (Giga) using a vacuum and centrifuge. Forour anion-exchange based Plasmid Purification Kits,a protocol can be accessed online at our Plasmid Resource Center, and is called 'Re-Purification of Plasmid DNA Prepared by Methods other than QIAGEN Tips'. The QIAprep Spin Miniprep Kit is intended for molecular biology applications. Thus, the plasmid DNA is highly suitable for applications that only allow small reaction volumes. Hose extender is not included. QIAprep Spin Miniprep Kit high-yield protocol for purification of up to 30 g plasmid DNA, Re-Purification of Plasmid DNA Prepared by Methods other than QIAGEN Tips, Isolation of plasmid DNA from mammalian cells using QIAprep kit, QIAGEN's nucleic acid purification technologies, Microcentrifuge or vacuum manifold; fully automatable using the QIAcube, Culture volume for low-copy plasmids/cosmids. Collection Microtubes and Caps. Try the Workflow Configurator. For purification of up to 10 g PCR products, 100 bp to 10 kb, For automated purification of ccfDNA from 1-96 samples using the QIAsymphony SP, For purification of up to 10 mg endotoxin-free advanced transfection-grade plasmid or cosmid DNA, For automated purification of high-quality DNA from 124 samples per run, For gel extraction/cleanup of up to 10 g DNA (70 bp to 10 kb) from enzymatic reactions, For automated purification of DNA from 196 samples on the QIAsymphony SP Instrument, For extraction of total cellular DNA from plant cells and tissues or fungi, or genomic DNA from plant cells, tissues and seeds, For purification of genomic DNA from formalin-fixed paraffin-embedded tissues, For automated high-throughput isolation of total DNA from blood, cells, and tissues, For ultrafast purification of up to 750 g transfection-grade plasmid or cosmid DNA, For purification of up to 20 g molecular biology grade plasmid DNA per well, For automated high-throughput purification of genomic DNA from fresh or frozen stool samples that are high in PCR inhibitors, For fast purification of up to 10 mg transfection-grade plasmid or cosmid DNA, For purification of up to 5 g PCR products (70 bp to 4 kb) in low elution volumes, For the isolation of microbial genomic DNA from all soil types, For removal of unincorporated dye terminators from 124 or 196 sequencing reactions, For the isolation of microbial DNA from stool and gut samples, For automated analysis of DNA fragments using QIAxcel instruments, For the isolation of genomic DNA from filtered water samples, including turbid water, For purification of up to 50 g high-quality plasmid DNA in 96-well format, For purification of up to 50 g transfection-grade plasmid DNA in 96-well format, For purification of archive-quality DNA from a wide variety of sample types, For the isolation of DNA from up to 384 soil samples in less than one day.