Plasmid DNA 정제
고품질 plasmid DNA는 클로닝 및 transfection부터 바이러스 패키징과 단백질 생산에 이르기까지 광범위한 분자생물학 기술에 필수적입니다. VectorBuilder의 plasmid DNA 정제 서비스는 기초연구 과학자 및 기업 고객의 요구사항을 충족하기 위해 ug에서 g까지 커스텀 스케일로 신속하고 비용 효율적인 생산을 제공합니다. 아래의 Research-grade, Research Plus-grade 및 Industrial-grade plasmid 외에도 임상 적용을 위한 GMP plasmid 제조도 제공합니다.
보다 나은 안전성 프로필, 제조 가능성 및 효능을 위해, antibiotic-free 및 supplement-free인 VectorBuilder의 독자적인 miniVec™ plasmid를 확인하세요.
중점 사항
Research Grade
- 클로닝 및 다양한 분자생물학 실험에 적합
- 신속한 소요시간과 비용 효율적
Research Plus Grade
- 세포 transfection, 바이러스 패키징, 단백질 발현, in vivo 연구 등 다양한 응용 분야에 적합
- 100 ug ~ 10 mg 범위의 스케일
- 높은 supercoil 비율 (80±10%) 및 낮은 endotoxin 레벨 (<0.05 EU/ug)
Industrial Grade
- 전임상 적용 및 대규모 다운스트림 생산에 적합
- 커스텀 생산 스케일 (100 ug ~ 1 g) 및 QC 분석
- 높은 supercoil 비율 (≥85%) 및 낮은 endotoxin 레벨 (<0.01 EU/ug)
- Animal-free 생산 가능
Order your desired scales with rapid turnaround.
서비스 상세
가격 및 소요시간
| 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 생산
VectorBuilder는 animal-product free plasmid DNA 정제 서비스를 제공하며, plasmid에 TSE (transmissible spongiform encephalopathies) 및 BSE (bovine spongiform encephalopathy)가 없음을 보장합니다. VectorBuilder는 배양 배지에 식물성 tryptone 대체물을 사용하고 DNA 정제를 위해 비효소적 방법을 사용합니다. 규제용 서류 제출을 지원하기 위한 TSE/BSE statement도 별도 요청시 가능합니다. 자세한 서비스 proposal을 위해 디자인 의뢰하기를 보내주세요.
배송 및 보관
Plasmid DNA는 1x TE buffer에 저장되어 있으며, 상온으로 배송됩니다. Plasmid DNA는 4°C에서 단기간 (2주 이하) 보관할 수 있습니다. 장기간 보관하는 경우 plasmid를 -20 °C 또는 -80 °C에 보관하세요. 반복적인 동결-해동은 피하시기 바랍니다.
고객 제공 시료
고객이 제공하는 벡터가 필요한 경우 시료 제출 가이드라인에 따라 plasmid를 준비하시고, 시료 샘플의 지연이나 손상을 방지하기 위해 가이드라인을 따라 주시기 바랍니다. 고객이 제공한 모든 시료는 VectorBuilder의 필수 QC를 거치며 각 항목에 대해 $50의 추가 요금이 발생할 수 있습니다. 고객이 제공한 시료가 QC를 통과할 때까지 생산이 시작되지 않을 수 있습니다.
기술적인 정보
품질 관리
| 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 | ||
| 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 | Not visible on agarose gel | N/A | ✓ | ✓ | |
| Fluorometric quantification | N/A | N/A | Optional | ||
| Host E. coli DNA | Not visible on agarose gel | 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)
|
정보 자료
주요 인용 논문
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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