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pDNA Synthesis
Custom synthesis of pDNA involves chemically synthesizing target DNA sequences into synthetic DNA molecules according to specific design requirements. BOC Sciences offers a variety of production tools to facilitate efficient pDNA production. Explore the process of plasmid DNA production from bacteria to final filtration and discover solutions to assist you at every step of the process.
What is pDNA?
Plasmid DNA (pDNA) is an important starting point for many genetic engineering studies, including the development of recombinant proteins, viral vectors and advanced biotherapeutics. pDNA can be replicated independently (free-form) from bacterial genomic DNA, a property that makes them ideally suited as vectors for genetic engineering.
Fig 1. Schematic of pDNA vectors and DNA minicircles. (H Sum et al., 2014)
pDNA Synthesis Services from BOC Sciences
pDNA Production
The target gene is inserted into the pDNA and then introduced into the host bacteria by electroporation. The host cell replicates during fermentation, replicating the plasmid and obtaining the desired sequence in high yield. Once the cells reach the desired density, the cells are harvested and the pDNA is extracted. The lysate is then clarified and the pDNA is purified using chromatography.
The types of pDNA production we can currently offer include the following.
| Research grade pDNA | Transfection grade pDNA | Quasi-medical pDNA |
| Research grade pDNA can be used for molecular cloning, targeted mutagenesis, bacterial transformation and other basic molecular biology experiments. | Transfection grade pDNA can be used for cell transfection, gene vaccine, gene therapy and other high-demand experiments. | For preclinical use in cell therapy, gene therapy, etc. |
Free Codon Optimization
BOC Sciences' biotechnology platform is able to optimize the sequences you need, according to your requirements, for any optimization host of your choice.
pDNA Manufacturing
In pDNA manufacturing production, the bacterial fluids are collected first, followed by alkaline lysis of the phage and finally removal of impurities, taking into account efficiency and amplification. pDNA manufacturing proceeds as follows.
Vector construction - Seed bank establishment - Fermentation - Phage collection - Solid-liquid separation of lysed cells - Clarification and concentration - Isolation and purification - Product quality control - Dispensing
Quality Control of pDNA
High-quality plasmid DNA is a key component of gene therapy production and is in high demand, requiring optimization of the production purification process to meet the quality requirements needed for therapeutic drug production. Quality control of plasmid DNA includes the following.
- Appearance by visual inspection
- Closed loop structure >85% (visual inspection)
- Sequence verification of the gene of interest
- Restriction analysis for larger sizes
Delivery Content
- High purity lyophilized plasmid DNA containing the target fragment and microorganisms containing the recombinant plasmid.
- Shipping documents, including standard sequence, sequencing results, sequence comparison documents, and shipping sample (COA) report.
Advantages of Our Services
- Ultra-low endotoxin levels:<0.1 EU/µg pDNA on selected scales (generally considered endotoxin-free)
- Scale-up: production from microgram to milligram scale
- Delivery: liquid or dry ice frozen
- Customized dispensing and tube labeling
BOC Sciences provides professional pDNA preparation and scale-up production services to help you complete your pDNA synthesis project efficiently. If you are interested in our services, please feel free to contact us.
Reference
- H Sum C, et al. Impact of DNA vector topology on non-viral gene therapeutic safety and efficacy[J]. Current gene therapy, 2014, 14(4): 309-329.
* Only for research. Not suitable for any diagnostic or therapeutic use.
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