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CleanCap® M6 IVT Kit, High Yield

SKU: K-7453
Need even higher protein expression than CleanCap M6 mRNA? Explore our combo CleanCap® IVT kit with ModTail™ kit

Description

TriLink’s CleanCap® M6 IVT Kit, High Yield is a comprehensive solution for mRNA synthesis by in vitro transcription (IVT) with CleanCap® co-transcriptional capping. The kit components are sufficient for 20 x 20 µL or 125 x 20 µL reactions depending on the kit size and following our recommended IVT protocol with co-transcriptional capping. When used with the pulse feed protocol, each 20 µL reaction typically produces 160–200 µg of capped mRNA, corresponding to approximately 3.2–4 mg per 20-reaction kit (K-7453-20) and 20–25 mg per 125-reaction kit (K-7453-125).

The kits are designed for cumulative gains in yield, quality, and expression. Key benefits of TriLink’s IVT kits compared to others with enzymatic or other co-transcriptional capping include:

  • Time and cost savings — Co-transcriptional capping protocol for high-yield mRNA synthesis with >95% capping efficiency in a single tube, reducing steps and hands-on time
  • High protein expression — Increase protein expression by ≥30% with CleanCap M6 versus its predecessors and enzymatic capping
  • Increased mRNA yield — Optimized CleanCap M6 IVT with a supplementary pulse-feed protocol for up to 10 mg/mL yield
  • Ultra-low double-stranded RNA (dsRNA) levels — Up to 85% dsRNA reduction using CleanScribe™ RNA Polymerase, in place of wild-type T7 RNA polymerase, in CleanCap M6 IVT
  • Enhanced mRNA performance — Achieve high protein expression and low inflammatory responses from mRNA synthesized with CleanCap M6, CleanScribe RNA polymerase, and N1-methylpseudouridine

The kit contains sufficient quantities of the following components to support the specified number of reactions:

  • CleanCap® Reagent M6
  • Adenosine-5'-Triphosphate
  • Cytidine-5'-Triphosphate
  • Guanosine-5'-Triphosphate
  • Uridine-5'-Triphosphate
  • N1-Methyl-Pseudouridine-5'-Triphosphate
  • M6 CleanScribe™ RNA Polymerase Mix
  • 10X M6 IVT Buffer
  • FLuc Control DNA Template

Important note:

  • This kit is specifically formulated & optimized for mRNA synthesis with CleanCap® M6 capping. It is not optimal for synthesis of saRNA or use with other cap analogs.
  • Some 5′ UTR sequences may lower mRNA yields using this CleanCap® M6 kit. Please review the protocol’s template design section or contact support@trilinkbiotech.com if you encounter 5′ UTR issues.

Product details
Catalog No. K-7453
Recommended Storage -15 to -20°C
Application In vitro Transcription, co-transcriptional capping, mRNA synthesis
Cap AG Start, Cap 1, CleanCap M6
Cap Analogs CleanCap M6
Product Grade RUO
Reaction Size 20 x 20 µL (K-7453-20), 125 x 20 µL (K-7453-125)

Technical documents

Product FAQs

We have observed the yield of CleanCap M6 IVT kit can be impacted by some 5' UTR sequences. Please refer to the CleanCap® M6 IVT kit protocol for details or contact support@trilinkbiotech.com for technical support if you are concerned about your UTR.

Optimization of the 5' and 3' untranslated regions (UTRs) depends on your specific goals and the target cell or tissue type for your mRNA.   To support this, our Officinae Bio services offer expert optimization services. Send us your sequence and UTRs, and we’ll perform codon optimization to help ensure optimal expression. Additionally, reviewing published literature can be a valuable starting point for identifying UTR sequences relevant to your application. We also recommend conducting screening experiments to evaluate translational efficiency. If you are using TriLink’s mRNA services, we have our own UTR sequences integrated in the DNA template as an option to insert your open reading frame (ORF) for a streamlined workflow.

The standard protocol of the kit typically results in 80-100 µ of RNA per 20 µL of reaction, and supplementing with its pulse feed protocol typically produces 160-200 µg of RNA per starting 20 µL of reaction. If your yield is lower, run a control reaction with the included FLuc control DNA template alongside your own template for troubleshooting. Also make sure 5′ UTR of your template is not an issue (review template design section of CleanCap® M6 IVT kit protocol). Please contact support@trilinkbiotech.com for further support.

Our IVT kits have a shelf life of at least one year from the date of manufacture. We also ensure that no kits are shipped if they have fewer than three months of remaining shelf life.

It is possible by adjusting the component amounts proportionally. Note that you may need to optimize reaction conditions to increase mRNA yield linearly while maintaining quality.

We do not recommend using CleanCap M6 IVT kit's buffer as a replacement for the restriction enzyme buffer. The kit buffer is low in pH (<2), rendering most restriction enzymes inactive.

The included buffer is specifically formulated for its corresponding kit and should not be used with another kit or cap for optimal results.

Using in vitro and in vivo experiments, we have consistently observed higher protein expression (≥30%) from mRNAs capped with CleanCap M6 versus CleanCap AG (3′ OMe) and enzymatic caps.  The increased protein expression is likely due in part to the N6-methylated adenosine on the CleanCap M6 analog impairing Dcp2-mediated decapping (Mandell & Ujita et al. (2025)).

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® M6 IVT Kit, High Yield, 20 x 20 µL rxns (sold as Each)
    SKU: K-7453-20
    $748.00
  • CleanCap M6 IVT Kit, High Yield, 125 x 20 µL rxns (sold as Each)
    SKU: K-7453-125
    $2,990.00
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