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Wizard MagneSil Tfx™ System
The Wizard MagneSil Tfx™ System provides a reliable method for the
rapid isolation of plasmid DNA in a multiwell format. The purified plasmid can be used directly for transfection, as well as for other standard molecular biology techniques.
A novel Endotoxin Removal Resin ensures high-quality plasmid DNA suitable for transfecting eukaryotic cells.
The use of MagneSil™ Paramagnetic Particles
(PMPs) for lysate clearing and DNA capture circumvents the need for cumbersome centrifugation or vacuum
manifolds, allowing DNA purification with the Wizard MagneSil Tfx™ System
to be completely automated.
Protocol Overview
The Wizard MagneSil Tfx™ System protocol is identical to the Wizard® MagneSil™ Plasmid Purification System with the addition of two steps designed to decrease contaminant levels and increase transfection efficiency. Lysate is treated with Endotoxin Removal Resin before the DNA is bound to MagneSil™ RED to remove endotoxin.
The 4/40 wash solution containing 40% isopropanol and 4M guanidine is used to wash the DNA-bound MagneSil™ RED PMPs to eliminate other contaminants. Through the use of these reagents, the resulting plasmid is of very high quality. A summary of the protocol is shown below.
Figure 1. Flow chart of transfection-grade plasmid DNA purification using the Wizard MagneSil Tfx™ System.
Features
- Purifies Transfection-grade DNA: Plasmid DNA purified using this system transfects a range of mammalian cell lines as well as or better than comparable competitive systems. This high quality DNA is also suitable for other molecular biology applications, including sequencing and restriction digestion.
- Effective Removal of Endotoxin: The Endotoxin Removal Resin supplied with the system cuts endotoxin carryover by as much as 95% over standard silica-based plasmid purifications systems. Lower endotoxin levels lead to better transfection efficiency.
- Saves You Time: Process a 96-well plate in 45 minutes with a robotic workstation. Multiple plates can be processed even faster with multiple magnets. Users save time and maximize throughput.
- Walk-away Automation: A validated method for Beckman Biomek® FX available
here. No manual intervention needed after pelleted cells are placed on the robot.
- Scalable-Whatever Your Throughput Needs: The scalable nature of
MagneSil™ Technology allows the researcher to use as little or as much paramagnetic particle suspension necessary to bind the amount of nucleic acid needed without incurring excess cost from wasted binding capacity. Elution volumes can be minimized to effectively purify and concentrate sample.
High Yields of Plasmid DNA
Many factors affect the final yield of plasmid DNA including; plasmid copy number, cell density and consistency of yield from sample-to-sample. The system has been tested with samples of varying cell densities to ensure consistent high yields of plasmid DNA.
Figure 2. Sample-to-sample yield of plasmid DNA purified using the Wizard MagneSil Tfx™ System.
The pGL3 Control Vector was purified from JM109 cells grown overnight in Terrific Broth (4.0
O.D.600 cell per well).
Transfection Grade DNA
Silica-based (sequencing grade) plasmid purification systems are routinely used for plasmid isolation due to their low-cost, speed and ease-of-use. However, DNA purified using these methods typically contains high levels of endotoxin. Anion exchange methods to isolate high quality, transfection-grade DNA can be time consuming and expensive. The Wizard MagneSil Tfx™ System offers a rapid, cost-effective method for purifying high quality, transfection-ready DNA. Plasmid DNA purified using this system transfects a range of mammalian cell lines as well as or better than, anion exchange methods. This high quality DNA is also suitable for other molecular biology applications.
Figure 3. A comparison of transfection efficiencies for different DNA purification systems with various cell lines. A variety of eukaryotic cell lines were transfected with
the pGL3 Control Vector purified using the Wizard MagneSil Tfx™ System, QIAGEN Turbo or QIAGEN Ultra systems. Transfection efficiency as a function of light output was compared for plasmid purified using silica membrane (QIAGEN Turbo) and anion exchange-based (QIAGEN Ultra) chemistries. Transfection efficiency was normalized as a percentage of anion exchange.
Effective Removal of Endotoxin
Endotoxin removal resin reduces endotoxin levels by 95% when using the Wizard MagneSil Tfx™ System. The purified DNA is suitable for
transfection into eukaryotic cells. Plasmid DNA purified using standard silica-based chemistries contains high amounts of endotoxin. The presence of endotoxin as a contaminant in plasmid DNA can result in reduced
transfection efficiency. For maximum consistency across multiple cell lines, it is critical to isolate plasmid DNA with low endotoxin levels.
Figure 4. Comparison of endotoxin levels for plasmid DNA purified using different methods. Plasmid DNA purified using the Wizard MagneSil Tfx™ System was compared with a number of other commercially available plasmid purification systems including a silica membrane-based system (QIAGEN Turbo) and an anion exchange system (QIAGEN Ultra).
Additional Information
Technical Bulletin
DNA and RNA Purification
Automated Methods
Promega Notes
View Online Catalog Information
Wizard MagneSil Tfx™ System
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