Endotoxin-free plasmid DNA purification using 96-well plates—NucleoBond 96 Xtra EF
NucleoBond 96 Xtra EF, the high-throughput kit of the NucleoBond Xtra product line, is designed for isolating high-quality, endotoxin-free plasmid DNA in a 96-well plate format. Depending on the culture conditions and growth medium, approximately 2–4 µg of plasmid DNA (from a 96-well plate culture) or up to 50 µg of plasmid DNA (from a glass tube incubation) can be isolated per well.
NucleoBond 96 Xtra EF, the high-throughput kit of the NucleoBond Xtra product line, is designed for isolating high-quality, endotoxin-free plasmid DNA in a 96-well plate format. Depending on the culture conditions and growth medium, approximately 2–4 µg of plasmid DNA (from a 96-well plate culture) or up to 50 µg of plasmid DNA (from a glass tube incubation) can be isolated per well. Growing cultures in rich media, such as Circlegrow or TB, is recommended. The isolated plasmid DNA is of highest purity and can be used for transfection, even for sensitive cell lines such as neuronal cells. The kit technology is based on anion-exchange chromatography using improved silica material with greater DNA binding capacity, based on proven NucleoBond anion-exchange group MAE (methylaminoethanol) technology.
Procedure
In the NucleoBond 96 Xtra EF procedure, E. coli cells are lysed using SDS/alkaline lysis. Cell lysates are clarified using a NucleoBond Filter Plate and then loaded onto the equilibrated NucleoBond Xtra EF Plate for plasmid binding. After subsequent washes with buffers to eliminate endotoxins and other contaminants, plasmid DNA is eluted in a high-salt buffer and precipitated with isopropanol. Precipitate separation is achieved using a specially designed NucleoBond Finalizer Plate, eliminating the need for a time-consuming centrifugation step. The kit can be used with vacuum or centrifugation processing. The NucleoBond Xtra EF Plate is used under gravity flow to achieve a specific flow rate needed for efficient binding to the anion-exchange group.
Required hardware for vacuum processing and centrifugation
Processing NucleoBond 96 Xtra EF under vacuum requires the NucleoVac 96 Vacuum Manifold. Centrifuge processing requires a microtiter plate centrifuge that is able to accommodate the NucleoBond Filter/Finalizer Plates stacked on a square-well block or a rack of tube strips, and that can reach accelerations of 5,600–6,000g (bucket height: 85 mm). For further information about suitable centrifuges, please contact Technical Support.
Overview
- Plasmid DNA with less than 0.1 EU/μg for transfection of highly sensitive cells
- Endotoxin removal by additional washing step
- Processing of up to 5 ml of bacterial culture
- NucleoBond Filter Plate for convenient filtration of bacterial lysates
- Processing under vacuum or by centrifugation
- Innovative MN Wash Plate minimizes risk of cross-contamination
- NucleoBond Finalizer Plate to speed up DNA precipitation
More Information
| Technology | Anion-exchange chromatography |
| Format | 96-well plates |
| Processing | Manual (vacuum or centrifugation) |
| Lysate clarification | 96-well filter plates |
| Starting material | 1–5 ml E. coli culture |
| Vector size | <15 kb <300 kb (without NucleoBond Finalizer Plate) |
| Typical yield | 2–4 μg (1.5 ml LB/TB in 96-well culture plates) 10–50 µg (5 ml LB/TB in glass tubes) |
| A260/A280 |
1.80–1.95 |
| Elution volume | 100–200 µl |
| Preparation time | 120 min/plate |
| Binding capacity | 50 µg |
Applications
- Manual isolation of high-copy plasmids
- Typical downstream applications: transfection, cloning, sequencing, PCR, restriction analysis
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.
Plasmid kit selection guide
Our plasmid DNA purification kits help you obtain reliable, high-quality plasmid DNA in minutes. We provide a wide range of formats from minipreps to gigapreps, as well as options for high-throughput processing. Specialized kits are also available for purifying plasmid DNA suitable for various downstream applications, such as transfection, sequencing, etc.
Selection guide Transfection-grade mini prepsTakara Bio USA, Inc.
United States/Canada: +1.800.662.2566 • Asia Pacific: +1.650.919.7300 • Europe: +33.(0)1.3904.6880 • Japan: +81.(0)77.565.6999
FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES. © 2025 Takara Bio Inc. All Rights Reserved. All trademarks are the property of Takara Bio Inc. or its affiliate(s) in the U.S. and/or other countries or their respective owners. Certain trademarks may not be registered in all jurisdictions. Additional product, intellectual property, and restricted use information is available at takarabio.com.
