baseScribe Cap1 IVT Kit

Product Information

Reliable One‑Step Synthesis of Translationally Competent Cap1‑Capped mRNA

The baseScribe Cap1 IVT Kit is an optimized in vitro transcription (IVT) system for the robust production of Cap1‑capped mRNA from DNA templates containing a T7 promoter. Designed for reliability and ease of use, the kit provides high yields of high‑quality mRNA in a single co‑transcriptional reaction, ready for downstream applications in vitro or in vivo.

Fig.1: The baseScribe Cap1 IVT Kit from baseclick is perfectly suited for co-transcriptional Cap1-capping. The high-quality mRNA is ready to use for translation.

The kit supports transcription from linearized plasmid DNA or PCR‑derived templates and enables reproducible generation of translationally competent mRNA without the need for post‑transcriptional capping steps.

Optimized Enzyme System for Reliable RNA Synthesis

The performance of the baseScribe Cap1 IVT Kit is based on the same optimized enzymatic architecture described in baseclick’s IVT whitepaper. Key features include:

• a high‑performance, GMP‑grade T7 RNA polymerase engineered for efficient transcription,
• pyrophosphatase to remove inhibitory pyrophosphate and prevent premature reaction stalling,
• and a carefully balanced transcription buffer designed to support sustained enzyme activity.
• RNase inhibitor and DNase I for template removal

This combination enables robust transcription efficiency and reproducible RNA quality across different templates and production batches. A linear DNA control template is included to allow evaluation of RNA yield and integrity.

High Yield from Minimal DNA Input

Under optimized conditions, the IVT system delivers 140–180 µg RNA from 1 µg of linearized plasmid DNA or 200–500 ng PCR‑derived template per 20 µL reaction.

Each kit supports 20 standard IVT reactions, allowing efficient production of Cap1‑capped RNA from limited DNA input.

The high yield and small reaction volume make the kit suitable for applications requiring reliable RNA production without extensive scale‑up.

Co‑Transcriptional Cap1-Capping for Efficient Translation

The baseScribe Cap1 IVT Kit enables co‑transcriptional incorporation of a Cap1 structure, generating RNA with a native‑like 5′-cap during transcription. This integrated approach eliminates the need for post‑transcriptional capping steps and streamlines RNA production workflows.

The resulting Cap1‑capped RNA is suitable for downstream applications requiring translationally competent RNA.

Demonstrated RNA Quality and Homogeneity

RNA produced using the optimized IVT system shows:

• high integrity and homogeneity,
• well‑defined transcript bands in agarose gel analysis,
• and minimal formation of truncated by‑products.

These characteristics are a direct result of optimized enzyme ratios, nucleotide purity, and controlled reaction conditions as demonstrated in the IVT whitepaper.

Compatibility with Modified Nucleotides

The baseScribe Cap1 IVT Kit is fully compatible with the incorporation of modified uridine analogs, including:

• pseudouridine (Ψ),
• N1‑methylpseudouridine (m¹Ψ),
• 5‑ethynyl uridine (EU)

Full replacement of uridine with Ψ or m¹Ψ does not reduce RNA yield, while incorporation of EU leads to an expected yield reduction of approximately 50%. This enables the generation of Cap1‑capped RNA tailored for stability, reduced immunogenicity, or downstream chemical functionalization.

Scalable and Reproducible Performance

The IVT reaction conditions have been optimized for reproducibility and scalability, enabling consistent performance across multiple reactions. Concentrated enzyme formulations contribute to enhanced storage stability and batch‑to‑batch consistency.

Key Features and Benefits

• Co‑transcriptional synthesis of Cap1‑capped RNA
• High‑performance T7‑based IVT system
• 140–180 µg RNA per reaction from 1 µg DNA
• GMP‑grade enzymes and nucleotides
• Pyrophosphatase‑supported reaction performance
• Compatible with modified and clickable nucleotides
• Reproducible and scalable IVT performance
• Control template for performance verification included

Applications

• mRNA‑based protein expression studies
• Functional mRNA analysis in vitro and in vivo
• Development and optimization of modified mRNAs
• Chemical biology and RNA labeling workflows

For research use only.

LITERATURE

Synthetic mRNAs with superior translation and stability properties, A. Grudzien‑Nogalska et al., Methods Mol Biol. 2013;969:55-72.

Incorporation of Pseudouridine Into mRNA Yields Superior Nonimmunogenic Vector With Increased Translational Capacity and Biological Stability, K. Karikó et al., 2008, Mol. Ther., Vol. 16, p. 1833–1840.

N1-methyl-pseudouridine in mRNA enhances translation through eIF2α-dependent and independent mechanisms by increasing ribosome density, I. Svitkin et al., Nucleic Acids Research, Volume 45, Issue 10, 2 June 2017, Pages 6023–6036.