In the preparation process of in vitro transcription (IVT) mRNA, linearized plasmid DNA is required as the transcription template for IVT with the help of T7 RNA polymerase. High-quality plasmid DNA is crucial for in vitro transcription.
Based on the mature plasmid preparation service platform, Yaohai Bio-Pharma can offer a linearized plasmid DNA preparation service of high purity and high standard to achieve efficient IVT transcription.
Services Details
| Process | Optional Service | Service Details | Delivery Period (Workday) |
| Circular plasmid preparation | Gene synthesis | Gene synthesis (third-party) | 7-10 |
| Plasmid amplification | Plasmid amplification | 2 |
| Plasmid extraction |
| Linearized plasmid preparation | Plasmid linearization and purification | Plasmid linearization | 1 |
| Linearization DNA purification |
| Plasmid DNA quality control | Concentration purity | Ultraviolet (UV) spectrophotometry | 1-2 |
| Plasmid conformation | Agarose gel electrophoresis (AGE) |
| Capillary electrophoresis (CE)-optional |
| Plasmid identify | Restriction enzyme identification/ AGE |
Our Features
Flexible selection of linearization methods
Optimization of DNA extraction and purification methods to achieve high recovery rates.
- Stringent Quality Control
Stringent process control standards, with a superhelical conformation rate of plasmid samples for research of over 80%.
High standard and high efficiency plasmid preparation and quality control services to meet experimental needs.
Case study
Taking Yaohai Bio-Pharma catalog mRNA_luciferase product as an example, the transcription template plasmid sample (research grade) has a superhelical ratio of more than 90%, a linearization ratio close to 100%, and a subsequent transcription ratio up to 1:200 (linearized plasmid DNA:mRNA).
The mRNA_luciferase obtained through the preparation of a linearized plasmid as a template is transfected into 293T cells, and the enzyme-substrate reaction activity is evaluated 24 h after transfection.
An obvious strong luciferase activity signal can be detected, i.e., the luciferase protein is expressed efficiently, suggesting the purity of the transcription template can fully satisfy the requirement of high-quality mRNA preparation.
Supercoiled Plasmid Ratio Assay
Validation of luciferase mRNA Expression In Vitro
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