CircRNA preparation requires linear RNA as precursor material (RNA precursor) for cyclization, where the RNA precursor is usually prepared using linearized plasmid DNA as a transcription template and transcribed in vitro with the help of T7 RNA polymerase.
High quality plasmid DNA is essential for in vitro transcription (IVT). Based on the mature plasmid preparation service platform, Yaohai Bio-Pharma can provide a high purity and standard linearized plasmid DNA (pDNA) preparation service to ensure the integrity of IVT products.
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’s product mCherry circRNA as an example, the supercoiled ratio of transcription template plasmid samples (research grade) is more than 70%, with a linearization rate of close to 100%.
mCherry circRNA is prepared with linearized plasmid as a template and transfected into 293T cells. High level of fluorescence expression (red fluorescence) is detected after 24 hours of transfection, which will be enhanced after 48 hours, and can still be detected on the 7th day and the 14th day after transfection.
mCherry protein is stably and efficiently expressed, and the transcriptional template purity can meet the requirement of circRNA drug product (DP) with high quality.
In vitro expression validation of mCherry circRNA
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