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CleanCap® M6 IVT Kit, High Yield

SKU: K-7453

Description

TriLink’s CleanCap® M6 IVT Kit, High Yield is a comprehensive solution for mRNA synthesis byin vitro transcription (IVT) with CleanCap® co-transcriptional capping. The kit components are sufficient for 25 x 100 µL or 125 x 20 µL reactions under the recommended protocol. When used with the pulse feed protocol, each kit is expected to yield 20-25 mg of capped mRNA.

Our kit’s key benefits 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 or 1 mg mRNA per 100-µL reaction from 5 µg linearized plasmid
  • 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 optional N1-methylpseudouridine

Kit components:

  • CleanCap® Reagent M6, 25 µmol
  • Adenosine-5'-Triphosphate, 100 µmol
  • Cytidine-5'-Triphosphate, 100 µmol
  • Guanosine-5'-Triphosphate, 100 µmol
  • Uridine-5'-Triphosphate, 100 µmol
  • N1-Methyl-Pseudouridine-5'-Triphosphate, 100 µmol
  • M6 CleanScribe™ RNA Polymerase Mix, 250 µL
  • 10X M6 IVT Buffer, 1 mL
  • FLuc Control Plasmid, 25 µg

Important note:

  • This kit is specifically formulated 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 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 25 x 100 µL or 125 x 20 µL

Technical documents

Product FAQs

They are stable for at least one year from the manufactured date.

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)).

We have observed the yield of CleanCap M6 IVT kit can be impacted by some 5' UTR sequences. Please 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 0.4-0.5 mg of RNA per 100 µL of reaction, and supplementing with its pulse feed protocol typically produces 0.8-1 mg of RNA per starting 100 µL of reaction. If your yield is lower, run a control reaction with the included FLuc control plasmid alongside your own template for troubleshooting. Also make sure 5′ UTR of your template is not an issue. Please contact support@trilinkbiotech.com for further support.

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.

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