Plasmid DNA Preparation
High-quality plasmid DNA is integral to a wide range of molecular biology techniques, from cloning and transfection to virus packaging and protein production. VectorBuilder's plasmid DNA preparation service offers rapid, cost-effective production on a customizable scale, from micrograms to grams, to meet the needs of basic research scientists and industrial customers. In addition to the Research-grade, Research Plus-grade and Industrial-grade plasmids below, we also offer GMP plasmid manufacturing for clinical applications.
For a better safety profile, manufacturability, and efficacy, check out our proprietary MiniVec™ plasmid for antibiotic-free and supplement-free preparation.
Highlights
Research Grade
- Suitable for cloning and various molecular biology experiments
- Cost effective with rapid turnaround
Research Plus Grade
- Suitable for various applications such as cell transfection, virus packaging, protein expression, and in vivo studies
- Scales ranging from 100 ug to 10 mg
- High supercoil ratio (80±10%) and low endotoxin level (<0.05 EU/ug)
Industrial Grade
- Suitable for preclinical applications and large-scale downstream production
- Highly customizable production scales (100 ug to 1 g) and QC assays
- Higher supercoil ratio (≥85%) and lower endotoxin level (<0.01 EU/ug)
- Animal-free production available
Order your desired scales with rapid turnaround.
Service Details
Price and turnaround
| Grade | Scale | Yield | Price (USD) | TAT | Application |
|---|---|---|---|---|---|
| Research | Mini | High copy: >10 ug (>200 ng/ul, 50 ul) | $39 | 1-2 days | Cloning and various molecular biology experiments |
| Medium and low copy: >5 ug (>100 ng/ul, 50 ul) | |||||
| Research Plus | Midi | High copy: >100 ug (>1 ug/ul, 100 ul) | $99 | 2-4 days | Various cell culture (e.g. transfection, virus packaging, protein expression, in vitro assays) and in vivo experiments |
| Medium and low copy: >50 ug (>500 ng/ul, 100 ul) | |||||
| Maxi | High copy: >300 ug (>1 ug/ul, 300 ul) | $149 | |||
| Medium and low copy: >150 ug (>500 ng/ul, 300 ul) | |||||
| Mega | High copy: >1 mg (>1 ug/ul, 1 ml) | $249 | |||
| Medium and low copy: >0.5 mg (>500 ng/ul, 1 ml) | |||||
| Giga | High copy: >10 mg (>1 ug/ul, 10 ml) | $799 | 5-7 days | ||
| Medium and low copy: >5 mg (>500 ng/ul, 10 ml) | |||||
| Industrial | 100 ug ~ 1 g | Please inquire | Preclinical experiments (e.g. efficacy and safety profile studies) and large-scale production of recombinant protein, virus, and IVT RNA for industrial applications | ||
Animal-free production
We offer animal-product free plasmid DNA preparation services, ensuring plasmids are free from transmissible spongiform encephalopathies (TSE) and bovine spongiform encephalopathy (BSE). We use a plant-based tryptone substitute in our culture media and non-enzymatic methods for DNA purification. A TSE/BSE statement is available to support regulatory submissions. Please send us a design request for a detailed service proposal.
Shipping and storage
Our plasmid DNA is stored in 1x TE buffer and shipped at ambient temperature. Plasmid DNA can be stored at 4°C for short term (≤2 weeks). For long-term storage, please put the plasmid at -20°C or -80°C. Avoid freeze-thaw cycles.
Customer-supplied materials
If customer-supplied vectors are required, please send us your vectors following the Materials Submission Guidelines to set up shipment to avoid any delay or damage of materials. All customer-supplied materials undergo mandatory QC by VectorBuilder which may incur a $50 surcharge for each item. Please note that production may not be initiated until customer-supplied materials pass QC.
Technical Information
Quality control
| Attribute | QC Assay | Research Grade | Research Plus | Industrial Grade | |
|---|---|---|---|---|---|
| General Properties | Appearance | Visual inspection | ✓ | ✓ | ✓ |
| Concentration | Nanodrop | ✓ | ✓ | ✓ | |
| Identity | Plasmid size | Restriction digestion | ✓ | ✓ | ✓ |
| Insert sequence | Sanger sequencing | Optional | Optional | Optional | |
| Full-length plasmid sequence | Sanger sequencing | Optional | Optional | Optional | |
| NGS | Optional | Optional | Optional | ||
| Third-generation sequencing | Optional | Optional | Optional | ||
| Purity | OD260/280 | Nanodrop | ✓ | ✓ | ✓ |
| Supercoiled plasmid ratio | Agarose gel electrophoresis | N/A | ✓ 80 ± 10% |
✓ ≥85% |
|
| Residual protein | BCA | N/A | N/A | Optional | |
| Host E. coli RNA | Agarose gel electrophoresis | N/A | ✓ | ✓ | |
| Fluorometric quantification | N/A | N/A | Optional | ||
| Host E. coli DNA | Agarose gel electrophoresis | N/A | ✓ | ✓ | |
| qPCR | N/A | N/A | Optional | ||
| Safety | Sterility | Direct inoculation | N/A | ✓ | ✓ |
| Endotoxin level | Kinetic chromogenic LAL assay | N/A | ✓ <0.05 EU/ug |
✓ <0.01 EU/ug |
|
| Phage detection | PCR | N/A | Only performed for Gigaprep | ✓ (when scale >10 mg) | |
| Document | - | - | COA (upon request) | COA (upon request) |
1. COA
2. TSE/BSE statement (upon request)
|
How to Order
Order both vector cloning and plasmid DNA preparation
Resources
Featured Citations
Lactic acid promotes PD-1 expression in regulatory T cells in highly glycolytic tumor microenvironments
Cancer Cell, 2022, doi: 10.1016/j.ccell.2022.01.001
Products used:
- Plasmid DNA Preparation
Abstract: The balance of programmed death-1 (PD-1)-expressing CD8+ T cells and regulatory T (Treg) cells in the tumor microenvironment (TME) determines the clinical efficacy of PD-1 blockade therapy through the competition of their reactivation. However, factors that determine this balance remain unknown. Here, we show that Treg cells gain higher PD-1 expression than effector T cells in highly glycolytic tumors, including MYC-amplified tumors and liver tumors. Under low-glucose environments via glucose consumption by tumor cells, Treg cells actively absorbed lactic acid (LA) through monocarboxylate transporter 1 (MCT1), promoting NFAT1 translocation into the nucleus, thereby enhancing the expression of PD-1, whereas PD-1 expression by effector T cells was dampened. PD-1 blockade invigorated the PD-1-expressing Treg cells, resulting in treatment failure. We propose that LA in the highly glycolytic TME is an active checkpoint for the function of Treg cells in the TME via upregulation of PD-1 expression.
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Glycine decarboxylase maintains mitochondrial protein lipoylation to support tumor growth
Dzmitry Mukha, et al.
Cell Metabolism, 2022, doi: 10.1016/j.cmet.2022.04.006
Products used:
- Plasmid DNA Preparation
Abstract: The folic acid cycle mediates the transfer of one-carbon (1C) units to support nucleotide biosynthesis. While the importance of serine as a mitochondrial and cytosolic donor of folate-mediated 1C units in cancer cells has been thoroughly investigated, a potential role of glycine oxidation remains unclear. We developed an approach for quantifying mitochondrial glycine cleavage system (GCS) flux by combining stable and radioactive isotope tracing with computational flux decomposition. We find high GCS flux in hepatocellular carcinoma (HCC), supporting nucleotide biosynthesis. Surprisingly, other than supplying 1C units, we found that GCS is important for maintaining protein lipoylation and mitochondrial activity. Genetic silencing of glycine decarboxylase inhibits the lipoylation and activity of pyruvate dehydrogenase and impairs tumor growth, suggesting a novel drug target for HCC. Considering the physiological role of liver glycine cleavage, our results support the notion that tissue of origin plays an important role in tumor-specific metabolic rewiring.
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Dominant mutations in MIEF1 affect mitochondrial dynamics and cause a singular late onset optic neuropathy
Majida Charif, et al.
Molecular Neurodegeneration, 2021, doi: 10.1186/s13024-021-00431-w
Products used:
- Plasmid DNA Preparation
Abstract: Inherited optic neuropathies are the most common mitochondrial diseases, leading to neurodegeneration involving the irreversible loss of retinal ganglion cells, optic nerve degeneration and central visual loss. Importantly, properly regulated mitochondrial dynamics are critical for maintaining cellular homeostasis, and are further regulated by MIEF1 (mitochondrial elongation factor 1) which encodes for MID51 (mitochondrial dynamics protein 51), an outer mitochondrial membrane protein that acts as an adaptor protein to regulate mitochondrial fission. However, dominant mutations in MIEF1 have not been previously linked to any human disease. Using targeted sequencing of genes involved in mitochondrial dynamics, we report the first heterozygous variants in MIEF1 linked to disease, which cause an unusual form of late-onset progressive optic neuropathy characterized by the initial loss of peripheral visual fields. Pathogenic MIEF1 variants linked to optic neuropathy do not disrupt MID51's localization to the outer mitochondrial membrane or its oligomerization, but rather, significantly disrupt mitochondrial network dynamics compared to wild-type MID51 in high spatial and temporal resolution confocal microscopy live imaging studies. Together, our study identifies dominant MIEF1 mutations as a cause for optic neuropathy and further highlights the important role of properly regulated mitochondrial dynamics in neurodegeneration.
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