Rapid desalting and concentration of plasmid DNA preps using a vacuum manifold—NucleoSnap Finisher Midi/Maxi kits
Eluates from anion-exchange plasmid preparations (such as NucleoBond midipreps and maxipreps) contain high salt concentrations. Macherey-Nagel has developed a new vacuum-driven accessory, the NucleoSnap Finisher Midi/Maxi kits, that allows you to desalt and concentrate plasmid DNA in a few minutes without any loss of yield or quality. These kits are the fastest way to desalt and concentrate DNA after your NucleoBond plasmid DNA preps.
Eluates from anion-exchange plasmid preparations (such as NucleoBond midipreps and maxipreps) contain high salt concentrations. Macherey-Nagel has developed a new vacuum-driven accessory, the NucleoSnap Finisher Midi/Maxi kits, that allows you to desalt and concentrate plasmid DNA in a few minutes without any loss of yield or quality. These kits are the fastest way to desalt and concentrate DNA after your NucleoBond plasmid DNA preps. NucleoSnap Finisher Midi/Maxi kits supersede the commonly used and time-consuming isopropanol precipitation and centrifugation steps by allowing rapid DNA precipitation and concentration of large volumes under vacuum.
Principle/procedure
DNA is precipitated by Precipitation Buffer FB and filtered through the specially designed matrix of the novel NucleoSnap Finisher Columns, followed by a washing step with ethanolic Wash Buffer A4 to remove salts and impurities.
To enable the use of a tabletop centrifuge, the NucleoSnap Finisher Column consists of an upper funnel component that can be separated from the lower mini spin column component by simply snapping off the funnel component. Then the mini spin column can be centrifuged in a standard tabletop centrifuge to remove residual ethanol.
Afterward, plasmid DNA can be eluted in the supplied endotoxin-free H2O-EF. The applied elution volume can be adjusted from 100 μl to 1.5 ml according to the expected amount of plasmid DNA, in order to ensure an optimal yield and concentration.
Figure 1. The NucleoSnap finisher midi/maxi procedure.
Application data
Figure 2. Ultra-fast plasmid DNA desalting and concentration using the NucleoSnap Finisher Maxi kit. NucleoBond Xtra Maxi kit eluates were precipitated using either the NucleoSnap Finisher Maxi kit or a conventional isopropanol precipitation. Six preparations can be processed in only seven minutes with the NucleoSnap Finisher Maxi kit, about 1/10th the time required for isopropanol precipitation.
Figure 3. 100% DNA recovery using the NucleoSnap Finisher Maxi kit. DNA yields in NucleoBond Xtra Maxi kit eluates were measured before and after precipitation with the NucleoSnap Finisher Maxi kit and isopropanol. With the NucleoSnap Finisher kits, 100% DNA recovery can be achieved, compared to 90% for isopropanol.
Overview
- No time-consuming isopropanol precipitation
- New column design (snap-off column) for vacuum processing of large sample volumes
- Process 12 samples in less than 10 minutes
- 100% plasmid DNA recovery
More Information
| Technology | Silica-membrane technology |
| Processing | Vacuum processing (e.g., using NucleoVac 24 Vacuum Manifold), or elution by centrifugation |
| Format | Snap-off column |
| Compatibility | Eluates from NucleoBond preparations: Xtra Midi (EF), Xtra Maxi (EF), PC 20, PC 100, PC 500 (EF) |
| Starting material | DNA eluates from e.g., anion-exchange purification kits |
| Vector size | <25 kb |
| Typical recovery | 90–100% |
| Elution volume | ≥100 µl |
| Preparation time | <10 min/12 preps |
| Binding capacity | 1.5 mg |
Applications
- Quick concentration and desalination of plasmid and cosmid DNA eluates obtained by anion-exchange chromatography
- Typical downstream applications: transfection, in vitro transcription, automated or manual sequencing, cloning, hybridization, and PCR
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
Isolate up to 700 µg of transfection-grade plasmid DNA in only 35 minutes
Using novel chemistry and a unique snap-off column design, NucleoSnap Plasmid Midi provides rapid purification of up to 700 µg of transfection-grade plasmid. The snap-off columns allow processing of lysate from large 50-ml culture volumes, and multiple samples can be rapidly processed by vacuum using our NucleoVac 24 Vacuum Manifold.
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