mRNA Co-transcription and Synthesis Kit, BP-07
Co-transcriptional kit for Cap1 mRNA synthesis.
Product overview
mRNA Co-transcription and Synthesis Kit, BP-07, supports co-transcriptional synthesis of capped mRNA from T7-promoter DNA templates. The workflow uses T7 RNA polymerase, NTP substrates, and cap analogs to generate 5'-capped RNA according to the instruction manual.
Features and applications
- Co-transcriptional capping enables mRNA transcription and capping in a single step;
- A 1 μg template input can yield 100–200 μg of mRNA;
- The kit provides Cap GAG, achieving a capping efficiency of ≥90% (up to >95%);
- The kit also provides N1-Me-pUTP, which can further reduce immunogenicity.
- The synthesized mRNA can be used directly for in vitro translation and cell transfection experiments.
- RNA and mRNA synthesis workflow development.
- Preparation of RNA products using the protocol and components described in the instruction manual.
Components
| Components | Cat. No. | Volume |
|---|---|---|
| ATP Solution(100 mM) | BP-AS-01 | 100 μL |
| GTP Solution(100 mM) | BP-AS-02 | 100 μL |
| CTP Solution(100 mM) | BP-AS-03 | 100 μL |
| UTP Solution(100 mM) | BP-AS-04 | 100 μL |
| N1-Me-pUTP Solution(100 mM) | BP-AS-07 | 100 μL |
| T7 RNA Polymerase | BP-E01 | 100 μL |
| RNase Inhibitor(40 U/μL) | BP-E02 | 100 μL |
| Inorganic Pyrophosphatase(0.1 U/μL) | BP-E03 | 100 μL |
| DNase I (2 U/μL) | BP-E04 | 100 μL |
| Cap GAG(100 mM) | BP-AS-38 | 100 μL |
| 10×Co-transcription Buffer | BP-AS-31 | 100 μL |
| Precipitation Reagent | BP-AS-10 | 1 mL |
| RNase-free ddH2O | BP-AS-11 | 1 mL |
Performance data
1 mRNA Co-transcription: Template Compatibility and Substrate Flexibility
Description: To evaluate the transcription performance of the Biori mRNA Co-transcription and Synthesis Kit using four DNA templates of varying sizes. Transcription reactions were performed with each template under both natural (UTP) and modified (N1-Me-pUTP) nucleotide conditions. The resulting RNA products were analyzed for yield and integrity.
Conclusion: The Biori mRNA Co-transcription and Synthesis Kit efficiently transcribed all four DNA templates, yielding RNA products with high integrity and robust yields under both natural (UTP) and modified (N1-Me-pUTP) nucleotide conditions, demonstrating its broad template compatibility and substrate flexibility.
2 Capping efficiency analysis of Cap1-mRNA prepared with the mRNA Co-transcription and Synthesis Kit
Description: Cap1-mRNA synthesized using the Biori mRNA Co-transcription and Synthesis Kit was processed with the mRNA Capping Efficiency Detection Pretreatment Kit (Cat: BP-05-10) and subsequently analyzed by LC-MS to determine the capping efficiency.
Conclusion: The Cap1-mRNA prepared using the Biori mRNA Co-transcription and Synthesis Kit achieved a capping efficiency of up to 97.3%, as determined by LC-MS analysis, demonstrating the kit's capability to produce highly capped mRNA transcripts.
3 eGFP expression of Cap1-mRNA in 293T cells
Description: Cap1-mRNA prepared using the Biori mRNA Co-transcription and Synthesis Kit was transfected into 293T cells to evaluate its translational expression efficiency. eGFP fluorescence was monitored to assess the expression level.
Conclusion: The Cap1-mRNA prepared using the Biori mRNA Co-transcription and Synthesis Kit was efficiently expressed in 293T cells, as demonstrated by robust eGFP fluorescence, confirming the kit's capability to produce translationally competent mRNA.
Shipping and storage
| Condition | Recommendation |
|---|---|
| Storage | Store at -20±5°C. |
| Shipping | Ship frozen on dry ice or with validated frozen cold-chain packaging that maintains the specified frozen storage condition. |
| Handling | Avoid repeated freeze-thaw cycles; mix thoroughly after thawing where applicable. |
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