Vaccine-grade plasmid DNA purification—NucleoBond RS
NucleoBond RS columns enable large-scale isolation of vaccine-grade, endotoxin-free plasmid DNA from bacterial cultures utilizing peristaltic pumps or FPLC systems. The NucleoBond RS workflow achieves endotoxin levels ≤0.01 EU/μg DNA, ensuring compliance for processes with high regulatory standards such as mRNA-based vaccine production. After two efficient washing steps that remove endotoxins and other contaminants, plasmid DNA is eluted, precipitated, and easily reconstituted in H2O-EF, TE-EF, or any other suitable endotoxin-free buffer for further use. In addition, RS columns are suitable for preparative scale RNA Clean-up from pre-purified RNA or IVT reactions.
NucleoBond RS columns enable large-scale isolation of vaccine-grade, endotoxin-free plasmid DNA from bacterial cultures utilizing peristaltic pumps or FPLC systems. The NucleoBond RS workflow achieves endotoxin levels ≤ 0.01 EU/μg DNA, ensuring compliance for processes with high regulatory standards such as mRNA-based vaccine production. After two efficient washing steps that remove endotoxins and other contaminants, plasmid DNA is eluted, precipitated, and easily reconstituted in H2O-EF, TE-EF, or any other suitable endotoxin-free buffer for further use. In addition, RS columns are suitable for preparative scale RNA clean-up from pre-purified RNA or IVT reactions.
For maximum flexibility, all components needed for plasmid purification or RNA clean-up using NucleoBond RS columns are offered as standalone products. For more detailed information regarding equipment and quantities of individual components, please refer to recommended reagents and equipment below.
Overview
A modular system for vaccine-grade plasmid purification or post-IVT RNA clean-up
- Achieve endotoxin levels ≤0.01 EU/μg DNA
- Process plasmids or post-IVT RNA using closed columns compatible with peristaltic pumps or FPLC
- Scale up from 10 mg to 200 mg bacterial culture (or 1 mg to 140 mg post-IVT RNA) with four column sizes
Recommended reagents for plasmid purification
Product | Size | Specification | Cat. # |
Buffer RES-EF | 1,000 mL | Resuspension buffer | 740386.1000 |
Buffer LYS-EF | 1,000 mL | Lysis buffer | 740387.1000 |
Buffer NEU-EF | 1,000 mL | Neutralization buffer | 740388.1000 |
Buffer EQ-EF | 1,000 mL 3,000 mL |
Equilibration buffer | 740380.1000 740380.3000 |
Buffer BMC-EF* | 1,000 mL 3,000 mL |
Binding mediating reconstitution buffer | 740382.1000 740382.3000 |
Buffer ENTO-EF | 1,000 mL 3,000 mL |
Endotoxin removal buffer | 740381.1000 740381.3000 |
Buffer WASH-EF | 1,000 mL | Wash buffer | 740392.1000 |
Buffer ELU-EF | 900 mL | Elution buffer | 740393.900 |
*Buffer BMC-EF is needed for the alternative RNAse-free NucleoBond RS plasmid purification workflow. Please refer to the NucleoBond RS Endotoxin-Free Plasmid DNA Purification User Manual for more information.
Recommended reagents for post-IVT RNA clean-up
Product | Size | Specification | Cat. # |
Buffer NRB | 1,000 mL 3,000 mL |
Binding buffer | 740371.1000 740371.3000 |
Buffer EQU | 1,000 mL | Equilibration buffer | 740317.1000 |
Buffer NRW | 1,000 mL 3,000 mL |
Wash buffer | 740372.1000 740372.3000 |
Buffer NRE | 1,000 mL 3,000 mL |
Elution buffer | 740373.1000 740373.3000 |
Additional recommended reagents and equipment
Product | Size | Specification | Cat. # |
Buffer TE-EF | 1,000 mL | Endotoxin-free TE buffer | 740797.1 |
Buffer H2O-EF | 1,000 mL | Endotoxin-free water | 740798.1 |
RNase A | 50 mg 100 mg |
Enzymatic degradation and removal of RNA | 740505.50 740505.100 |
NucleoBond Bottle Top Filter Type 2 | 5 Filters | Lysate filtration | 740553.5 |
NucleoVac 96 Vacuum Regulator | 1 piece | Vacuum manifold for controlling vacuum | 740641 |
More Information
Specifications of plasmid DNA isolated using NucleoBond RS columns
Parameter | Method | Criterion |
Integrity | Agarose gel electrophoresis/fragment analyzer | ≥90% supercoiled form |
Purity | UV spectrum | A260/A280 1.8–2.0 A260/A230 1.9–2.3 |
Endotoxin levels | LAL/rFC | ≤0.01 EU/μg DNA |
Host DNA | gDNA PCR | ≤1% |
Host RNA | Agarose gel electrophoresis Fluorometric quantification |
Not visible at 200 ng DNA ≤2% |
Host proteins | BCA | ≤1% |
Applications
- Large-scale, endotoxin-free plasmid purification
- Preparation of vaccine-grade plasmid DNA
- Post-IVT RNA clean-up
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.
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