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CleanCap® AU saRNA CleanScript™ IVT Kit

SKU: K-7114
Make your own high-quality saRNA in-house for less than sourcing the components yourself.

Description

TriLink’s CleanCap® AU saRNA CleanScript™ IVT Kit provides essential components for self-amplifying RNA (saRNA) or self-replicating RNA (srRNA) synthesis using co-transcriptional capping by in vitro transcription (IVT). The kit components are sufficient to run 20 × 20 µL reactions following our recommended IVT protocol. It includes a cap analog, modified and unmodified nucleotides, IVT enzymes, a reaction buffer, and a control DNA template. Each 20 µL reaction is expected to yield up to 150–190 µg of capped saRNA (up to 7.5–9.5 mg/mL, crude yield following LiCl purification) — approximately 3.0–3.8 mg per 20-reaction kit (K-7114-20).

As the first and only dedicated saRNA IVT kit, it is designed for cumulative gains in yield, quality, and expression. Key benefits include:

  • Time and cost savings — Co-transcriptional capping protocol with all necessary reagents for a single-tube reaction providing high yield, fewer steps, and less handling.
  • High capping efficiency — Protocol with CleanCap® AU cap analog to produce an optimal cap 1 with over 95% capping efficiency.
  • Increased saRNA yield — Buffer and kit components formulated using TriLink’s proprietary CleanScript® IVT method to yield up to 150–190 µg capped saRNA per 20 µL reaction from 2.4 µg linear plasmid.
  • Ultra-low double-stranded RNA (dsRNA) — Up to 85% dsRNA reduction (depending on modifications) using CleanScribe™ RNA Polymerase in place of wild-type T7 RNA polymerase in combination with the CleanScript IVT method.
  • Enhanced saRNA performance — Robust protein expression and low inflammatory responses from saRNA synthesized with CleanCap AU, CleanScribe RNA polymerase, and 5-methylcytidine (5mC).


Why the CleanCap saRNA IVT kit vs. sourcing the components yourself

Sourcing and optimizing the parts yourself means weeks of tuning Mg²⁺, temperature and buffer, failed runs, and no lot-to-lot guarantee. The kit is a co-optimized, saRNA-specific IVT system developed and validated by TriLink and lot verified on a 9.4 kb control, so you can produce high-quality saRNA on the first run without rebuilding the system in-house.

Route Time to data Optimization required Relative cost
CleanCap® AU Kit Same day None 1× (Reference)
Individual reagents Weeks High ≈3×
Outsourced synthesis Weeks–months None ≈8–10×

Bottom line: Once optimization time is factored in, the do-it-yourself approach can cost roughly 3× more than the kit. The CleanCap® AU saRNA IVT Kit delivers a pre-optimized workflow with kit-exclusive AU CleanScribe™ Mix and CleanScript™ IVT Buffer, helping you skip weeks of optimizing reaction conditions and get to high-quality saRNA faster.

The kit gives you the kit-only AU CleanScribe™ Mix, a saRNA-tuned CleanScript™ IVT buffer, and a pre-validated protocol so you spend less time & cost troubleshooting IVT and more time on the research and biology that matters to you.

The kit contains sufficient quantities of the following components to support 20 × 20 µL reactions:

  • CleanCap® Reagent AU
  • Adenosine-5′-Triphosphate (ATP)
  • Cytidine-5′-Triphosphate (CTP)
  • Guanosine-5′-Triphosphate (GTP)
  • Uridine-5′-Triphosphate (UTP)
  • 5-Methylcytidine-5′-Triphosphate (5mC)
  • AU CleanScribe™ RNA Polymerase Mix
  • 10X AU CleanScript™ IVT Buffer
  • saRNA FLuc Control DNA Template (linearized)

Important note:

  • This kit is specifically formulated and optimized for in vitro saRNA synthesis with CleanCap® AU capping. It is not optimal for standard mRNA synthesis or use with other cap analogs.
  • Users must design and source their own IVT template. PCR templates have not been validated for saRNA (see FAQ); use a fully linearized plasmid. Refer to the protocol’s[PK2.1] template design section or contact [email protected].

Product details
Catalog No K-7114
Recommended Storage -15 to -20 °C
Application In vitro transcription; co-transcriptional capping; self-amplifying RNA (saRNA / srRNA) synthesis
Cap AU Start, Cap 1, CleanCap AU
Cap Analogs CleanCap AU
Reaction Size 20 × 20 µL
Incubation 31 °C, 5 hours

Technical documents

Product FAQs

CleanCap AU preserves the authentic 5′ alphavirus end required for saRNA replication and provides an optimal Cap 1 structure with over 95% capping efficiency. AU-start saRNA templates require CleanCap AU; caps used for mRNA (e.g., AG or GG) are not suitable.

TriLink’s proprietary in vitro transcription method to produce high-quality, high-yield RNA while minimizing dsRNA. The kit’s reaction buffer is formulated on the CleanScript method and further optimized for long saRNA templates.

Yes. 5mC can be used in place of CTP to potentially suppress the interferon response and increase potency; replace CTP with the same concentration of 5mC.

A novel enzyme that substitutes wild-type T7 RNA polymerase in IVT and reduces dsRNA formation by up to 85%. The AU CleanScribe™ RNA Polymerase Mix also contains RNase inhibitor and inorganic pyrophosphatase.

DNase treatment is optional but recommended to remove template DNA. Use the one-pot DNase protocol in the manual or purify then DNase-treat.

The kit provides all necessary reagents sufficient for 20 × 20 µL reactions (K-7114-20).

Lithium chloride precipitation, spin columns (e.g., QIAGEN RNeasy), or oligo(dT). Oligo(dT) can yield higher-integrity saRNA but may be less efficient for VEE saRNA and require optimization.

A typical AU CleanScript™ IVT reaction yields 150–190 µg of saRNA per 20 µL reaction (up to 7.5–9.5 mg/mL, crude following LiCl purification) using unmodified NTPs or 5mC.

The template must contain the correct T7 promoter sequence (5′ TAATACGACTCACTATA 3′) followed by an initiating sequence of 5′ ATG 3′ (recommended) or 5′ ATA 3′ for proper use with CleanCap AU.

To verify saRNA transcription and aid troubleshooting. Following the recommended protocol it yields 150–190 µg of 9.4 kb saRNA per 20 µL reaction. 12 µg of linearized control template (sufficient for 5 × 20 µL reactions; 26A poly(A) tail) is provided.

The reaction conditions have been tested with templates up to ~18 kb, resulting in ~16 kb saRNA transcripts.

A 5-hour incubation at 31 °C, using a thermal cycler with a heated lid or a dry-air incubator. The 31 °C temperature is optimized to promote saRNA integrity.

Yes, by adjusting the component amounts proportionally.

No, the plasmid DNA must be fully linearized. Residual circular plasmid will produce unwanted extended RNA byproducts that can affect your results.

Yes, PCR products can be used directly, though using silica-membrane-purified PCR products typically gives better results. Please ensure it contains the correct T7 promoter sequence and the start site as described in the template design section. The final concentration should be 0.025 ug/uL for PCR templates (compared to 0.05 ug/uL for plasmid templates).

Certificate of analysis

Intellectual property

Products are for research use only, not for use in diagnostic or therapeutic procedures or for use in humans. Products are not for resale without express written permission from TriLink No license under any patent or other intellectual property right of TriLink or its licensors is granted or implied by the purchase unless otherwise provided in writing.

CleanCap capping technology

For Research Use Only. Not for use in humans. Not for use in diagnostic or therapeutic purposes. For additional licensing restrictions, please see the license agreement at trilinkbiotech.com/cleancap-research-license. Patents and patent pending, see trilinkbiotech.com/legal-notices.

  • CleanCap® AU saRNA CleanScript™ IVT Kit, 20 x 20 µL rxns (sold as Each)
    SKU: K-7114-20
    $848.00
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