Streamline Your RNA Synthesis and Capping While Enhancing Your Results Significantly

Get all-in-one IVT kits to produce high-yield, low-dsRNA capped RNAs in one step 

TriLink’s CleanCap® IVT kits include all the necessary components to produce mRNAs by in vitro transcription (IVT) and CleanCap® co-transcriptional capping.

Experience the difference with our IVT kits which incorporate TriLink’s innovative RNA technologies for:

  • 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
  • Up to 2x higher IVT yield than standard IVT protocols
  • 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

Which IVT Kit is right for you?

The choice depends on your desired protein expression level, downstream applications, and IVT experience.

  • CleanCap® AG (3′ OMe) CleanScript™ IVT Kit: It is a versatile kit that requires minimal optimization to synthesize high-yield, low-dsRNA capped mRNA. CleanCap® AG (3′ OMe) cap analog has been shown to provide higher protein expression than the original CleanCap® AG cap analog and is used in commercially approved mRNA vaccines.
  • CleanCap® M6 IVT Kit, High Yield: It includes our newest cap analog, CleanCap® M6, for those seeking higher protein expression than with enzymatic caps and previous CleanCap® AG analogs. Users may need to review their 5’-UTR sequence compatibility to achieve expected yields of up to 5 mg/mL using the standard protocol or 10 mg/mL using the supplementary pulse-feed protocol.

 

  CleanCap® AG (3′ OMe) CleanScript™ IVT Kit
  CleanCap® M6 IVT Kit, High Yield
Description CleanCap® Reagent AG (3′ OMe) is designed for the co-transcriptional capping of mRNAs to produce an optimal Cap 1. It includes the 5′ N7-methyl-3′-O-methylguanosine commonly found in mRNA capped using anti-reverse cap analog. Cap-1 mRNAs produced by CleanCap AG (3′ OMe) have superior in vivo activity compared to Cap-0 mRNAs produced by legacy co-transcriptional capping methods  

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)).

Cap

2’ O-methylation on the first base (Cap 1)
3’ O-methylation on m7G

AG Start

 

2’ O-methylation on the first base (Cap 1)
3’ O-methylation on m7G
m6A modification

AG Start

Considerations Versatile   Some 5′ UTR sequences may lower mRNA yields
Kit Component
  • CleanCap® Reagent AG (3' OMe), 10 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
  • AG CleanScribe™ RNA Polymerase Mix, 250 µL
  • 10X AG CleanScript™ IVT Buffer, 1 mL
  • FLuc Control Plasmid, 25 µg
 
  • 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
Expected  yield
(from 100 µL rnx)
 0.8-1 mg    0.8-1 mg
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. 
Reaction Size    
 25 rxns (100 µL scale) or125 rxns (20 µL scale)    25 rxns (100 µL scale) or 125 rxns (20 µL scale)
Catalog number  K-7413    K-7453-25

 

NOTE: The cap analog, buffer, and polymerase mix are optimized to each kit and should not be interchanged.

Can I use TriLink’s kit protocol to make uncapped mRNA?

With TriLink’s CleanCap AG (3′ OMe) and CleanCap M6 IVT kits for mRNA, you may omit the cap in their published protocol to obtain uncapped RNA. A similar yield is expected. However, if your template is 'AG start', there will be more heterogeneity at the 5' end, for example a mix of A and G. Depending on the required application of the uncapped mRNA, phosphatase treatment can be used to remove the 5' triphosphate if required.

 

Diana Perrier, Marketing Manager

Marketing Team at Tebubio

"Researchers across Europe are already moving to the CleanCap IVT Kit for faster, more reliable mRNA synthesis. With higher yields, cleaner transcripts, and a dramatically simplified protocol, it’s the smartest choice for modern mRNA production."


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  • References

    Article content created by Tebubio using courtesy materials provided by TriLink Biotechnologies.

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