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Cap Analogs for mRNA Synthesis & IVT Workflows
Explore cap analogs for mRNA synthesis, including Cap 0, Cap 1, ARCA, and chemical capping reagents for reliable IVT performance.
Cap analogs are a foundational component of mRNA synthesis, directly influencing mRNA stability, translation efficiency, and overall IVT performance. As mRNA-based technologies continue to advance, understanding how different cap analogs function—and how capping strategies affect downstream expression—has become increasingly important for reliable and reproducible mRNA production. We provides a clear overview of cap analogs used in IVT mRNA synthesis, explaining the role of 5' capping, comparing co-transcriptional and post-transcriptional capping approaches, and outlining why careful cap analog selection is essential for achieving high-quality mRNA and consistent protein expression outcomes.
Cap analogs are essential components in mRNA synthesis workflows, providing the chemical basis for introducing a functional 5' cap structure during in vitro transcription (IVT). By emulating the natural cap found on eukaryotic mRNA, cap analogs play a central role in protecting mRNA from degradation and enabling efficient translation. Modern cap analog designs address key challenges in IVT, including incorporation efficiency, cap orientation, immune recognition, and scalability, making them a foundational element of high-quality mRNA production.
The 5' cap is a modified guanosine structure located at the 5' end of mRNA and is indispensable for proper mRNA function. In IVT-derived mRNA, the cap protects transcripts from 5'–3' exonuclease-mediated degradation and facilitates interaction with cap-binding proteins that initiate translation. Efficient 5' capping directly supports mRNA stability, ribosome recruitment, and protein expression, and is therefore a critical quality attribute in any mRNA synthesis process.
Two primary strategies are used to introduce the 5' cap in mRNA synthesis: co-transcriptional capping and post-transcriptional capping. Co-transcriptional capping incorporates the cap structure directly during IVT, combining transcription and capping into a single step and offering advantages in workflow simplicity, scalability, and reproducibility. Post-transcriptional capping, by contrast, relies on enzymatic modification after RNA synthesis, allowing precise cap formation at the cost of additional processing steps and operational complexity. The choice between these approaches depends on process requirements, performance goals, and manufacturing considerations.
Cap analog selection has a direct and measurable impact on mRNA stability, translation efficiency, and overall IVT performance. Variations in cap structure, methylation pattern, and orientation control can influence how effectively mRNA is translated and how consistently it performs across batches. Some cap analogs are designed to prevent reverse incorporation, while others more closely mimic native mRNA to improve biological relevance. Selecting the appropriate cap analog ensures reproducible results, higher protein expression, and reliable mRNA quality throughout research and production workflows.
Our cap analog portfolio is designed to support a wide range of mRNA synthesis and in vitro transcription (IVT) workflows, addressing key performance factors such as capping efficiency, cap orientation, translation output, and process scalability. By offering multiple cap structures and capping strategies, we enable researchers and process developers to select the most appropriate solution based on their application requirements and workflow complexity. All products are manufactured with strict quality controls to ensure reliable incorporation and consistent IVT performance.
We provide a comprehensive selection of Cap 0, Cap 1, and Cap 2 analogs to accommodate different biological and technical requirements. Cap 0 analogs are widely used in basic mRNA research and early-stage studies where robust translation performance and workflow simplicity are priorities. Cap 1 analogs, which include an additional 2'-O-methyl modification on the first transcribed nucleotide, more closely resemble native eukaryotic mRNA and are commonly chosen for applications requiring improved translation efficiency and reduced immune recognition. Cap 2 analogs are available for specialized use cases that demand further structural modification of the cap.
Our portfolio includes anti-reverse cap analogs (ARCA) designed to prevent reverse incorporation during IVT. By controlling cap orientation, ARCA increases the proportion of correctly capped mRNA molecules, leading to more efficient recognition by translation initiation factors and improved protein expression. ARCA is particularly valuable in workflows where maximizing translation efficiency and minimizing variability are critical objectives.
In addition to traditional cap analogs, we offer chemical capping reagents for co-transcriptional capping during IVT mRNA synthesis. These reagents enable transcription and capping to occur in a single step, simplifying workflows and supporting high capping efficiency. Chemical capping systems are well suited for applications that prioritize process simplicity, scalability, and reproducibility, especially in mRNA production workflows requiring consistent performance across batches.
Our cap analogs are available in both research-grade and GMP-grade formats, allowing customers to align material selection with their development stage and quality requirements. Research-grade products support exploratory studies, assay development, and process optimization, while GMP-grade materials are manufactured under enhanced quality systems with comprehensive documentation to support regulated mRNA production environments.
Cap analogs are used across a broad range of mRNA synthesis and IVT workflows, where proper 5' capping is essential for achieving stable, functional, and efficiently translated mRNA. The specific cap analog and capping strategy selected can significantly influence translation efficiency, reproducibility, and process robustness. As a result, cap analogs play a central role not only in basic mRNA research, but also in advanced process development and scaled mRNA production.
In IVT workflows, cap analogs are incorporated during transcription to generate a functional 5' cap structure on newly synthesized mRNA. Proper capping improves mRNA stability, yield, and consistency, supporting reliable downstream processing and analysis. Cap analogs are therefore a foundational component of IVT mRNA production for a wide range of experimental and manufacturing-oriented workflows.
Cap analogs are critical for producing mRNA that can be efficiently translated in cell-free and cell-based expression systems. By enabling effective ribosome recruitment and translation initiation, correctly capped mRNA supports robust protein expression, making cap analogs essential for applications such as reporter assays, functional screening, and pathway analysis. Differences in cap structure and orientation can have a direct impact on observed expression levels.
During process development, cap analogs are evaluated and optimized to improve capping efficiency, translation performance, and reproducibility. Comparing different cap structures and capping strategies allows researchers to identify conditions that deliver consistent mRNA quality while minimizing variability. Cap analog selection is often a key optimization parameter in refining IVT workflows.
For workflows that involve increased reaction volumes or repeated production cycles, cap analogs must perform reliably under scaled conditions. High-quality cap analogs and chemical capping systems are commonly used to support scalable and reproducible mRNA production, where consistent performance, workflow simplicity, and supply reliability are critical to maintaining mRNA quality across batches.
The performance of cap analogs in mRNA synthesis and IVT workflows depends not only on chemical design, but also on the rigor of manufacturing and quality control processes. To support reliable mRNA production, our cap analogs are manufactured under strict quality and process control standards, ensuring consistent performance, reproducibility, and long-term supply reliability across research and production environments.
All cap analogs are produced to achieve high chemical purity with carefully controlled impurity profiles. Advanced analytical methods are used to verify chemical identity and structural integrity, helping ensure efficient cap incorporation during IVT and minimizing the risk of incomplete or defective capping. High purity directly contributes to improved mRNA stability and translation efficiency.
Consistent IVT performance requires materials with minimal lot-to-lot variability. Our manufacturing processes are designed to ensure batch-to-batch consistency, with each production lot subjected to standardized quality control testing. This reproducibility supports stable capping efficiency, predictable mRNA yield, and reliable translation performance across repeated IVT runs.
Cap analogs are manufactured using controlled and well-defined processes, with comprehensive quality documentation available to support integration into regulated or quality-driven workflows. Clear lot traceability, standardized testing protocols, and detailed product specifications help customers maintain control over their mRNA synthesis processes and ensure compliance with internal quality requirements.
In addition to product quality, supply continuity is essential for mRNA projects that scale over time. Our manufacturing capabilities are designed to support reliable and scalable supply of cap analogs, ensuring consistent availability as production demands increase. This reliability enables long-term planning and uninterrupted mRNA synthesis operations.
Selecting a cap analog supplier is a critical decision that directly affects mRNA quality, IVT reproducibility, and downstream translation performance. Our cap analogs are developed with a strong emphasis on chemical expertise, manufacturing control, and practical workflow compatibility, helping customers achieve consistent and reliable results across a wide range of mRNA synthesis applications.
Choosing the right cap analog is a foundational step in achieving stable mRNA, efficient translation, and reproducible IVT performance. Whether you are setting up a new mRNA synthesis workflow or optimizing an existing process, our comprehensive cap analog portfolio and technical expertise can help you identify the most suitable solution for your application. Contact us today to request technical information, discuss your requirements, or receive a customized quotation.
Cap analogs are used to introduce a functional 5′ cap during IVT mRNA synthesis, improving mRNA stability and translation efficiency.
Cap 1 analogs include an additional 2′-O-methyl modification that more closely mimics native mRNA and often supports improved translation.
Incorrectly oriented caps cannot be recognized by translation initiation factors, reducing protein expression efficiency.
ARCA is recommended when maximizing translation efficiency and reducing variability caused by reverse cap incorporation are important.
Most cap analogs are designed to be compatible with commonly used RNA polymerases and IVT workflows.
