Pre-treatment Kit for mRNA Poly A Distribution Detection, BP-11
Prepares mRNA samples for poly(A) distribution analysis.
Product overview
Poly A tail length is a critical quality attribute for mRNA vaccines and therapeutics, constituting essential validation data required for the regulatory submission of mRNA-based products. LC-MS (liquid chromatography-mass spectrometry) has emerged as a primary detection method due to its ability to precisely determine poly A tail length and heterogeneity, making it suitable for process development and quality control. The Pre-treatment Kit for mRNA Poly A Distribution Detection developed by BIORI is designed to efficiently and rapidly obtain the poly A tail of mRNA, accommodating sequences with different characteristics and various types of poly A tail designs. It offers product advantages such as high-efficiency purification, simple operation, high sensitivity and specificity, as well as safety and reliability.
Features and applications
- Efficiently and rapidly obtains the poly A tail of mRNA, accommodating sequences with different characteristics and various types of poly A tail designs.
- High-efficiency purification with magnetic beads and simple operation.
- Recommended digestion system: 50 μg of RNA in 50 μL with T Reagent and 10×T Buffer, digested at 37°C for 2 hours.
- For sample preparation prior to the detection of poly(A) distribution in capped mRNA.
Components
| Components | Volume | Storage | |
|---|---|---|---|
| Box 1 | T Reagent | 60 μL | -20±5°C |
| 10×T Buffer | 120 μL | ||
| Box 2 | Magnetic Beads | 1 mL | 2-8°C |
| Buffer I | 6 mL | ||
| Buffer II | 20 mL | ||
| EB Buffer | 1 mL×2 |
Performance data
1 Functional test of the kit
Description: The treated samples after enzymatic digestion and the recovered samples were verified using denaturing electrophoresis.
Conclusion: After digestion with RNase T1 using the kit, there were still considerable impurities. However, following purification and recovery, the purity of the Poly(A) tail band was significantly improved.
2 LC-MS analysis for determining the distribution of Poly(A)
Description: The recovered samples were subjected to LC-MS analysis to determine the distribution of Poly(A).
Conclusion: The ion current within the target peak exhibited a trend toward normal distribution. Through deconvolution and spectral analysis, a standard normal or near-normal distribution of the Poly(A) tail lengths could be obtained.
Table 1 Recommended reaction system.
| Components | Volume |
|---|---|
| T Reagent | 5 μL |
| RNA | 50 μg |
| 10×T Buffer | 5 μL |
| RNase-free ddH2O | to 50 μL |
Shipping and storage
| Condition | Recommendation |
|---|---|
| Storage | Box 1 is stored at -20±5°C and Box 2 at 2-8°C. Do not freeze-thaw the magnetic beads. |
| Shipping | Ship with split cold-chain packaging: frozen components at -20°C on dry ice and refrigerated components at 2-8°C with cold packs. |
| Handling | Avoid repeated freeze-thaw cycles; mix thoroughly after thawing where applicable. |
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