arrow_back Back

CleanCap® AG (3′ OMe) CleanScript™ IVT Kit - (K-7413)

SKU: K-7413

Description

TriLink’s CleanCap® AG (3′ OMe) CleanScript™ IVT Kit provides essential components for mRNA synthesis using co-transcriptional capping by in vitro transcription (IVT). The kit components are sufficient to run 25 x 100 µL or 125 x 20 µL reactions reactions following our recommended IVT protocol with co-transcriptional capping. It includes a cap analog, modified and unmodified nucleotides, IVT enzymes, a reaction buffer, and a control DNA template. Each kit is expected to yield 20-25 mg of capped mRNA.

The kit is designed for:

  • 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 AG (3′ OMe) cap analog to produce an optimal cap 1 with over 95% capping efficiency
  • Increased mRNA yield — Buffer and kit components formulated using TriLink’s proprietary CleanScript IVT method for up to 10 mg/mL yield (or up to 1 mg mRNA from 5 µg linear plasmid) per 100 µL reaction
  • 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 combination with the CleanScript method (instead of standard IVT)
  • Enhanced mRNA performance — Robust protein expression and low inflammatory responses from mRNA synthesized with CleanCap AG (3′ OMe), CleanScript method, CleanScribe RNA polymerase, and optional N1-methylpseudouridine

The kit includes:

  • 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

Product details
Recommended Storage -15 to -25°C
Application In vitro Transcription, co-transcriptional capping, mRNA synthesis
Cap AG Start, Cap 1, CleanCap AG (3′ OMe)
Cap Analogs CleanCap AG (3' OMe)
Reaction Size 25 rxns (100 µL scale), 125 rxns (20 µL scale)

Technical documents

Product FAQs

The reaction conditions have been tested with templates up to 6 kb in length.

CleanScribe RNA Polymerase is a novel enzyme designed to substitute wild-type T7 RNA polymerase in IVT and reduce dsRNA formation by up to 85%.

The kit contains an IVT buffer for co-transcriptional capping with the supplied CleanCap analog. The buffer is proprietary and formulated based on TriLink's CleanScript IVT method to increase yield and lower dsRNA formation.

The kit provides all necessary reagents sufficient for 25 x 100 uL reactions or 125 x 20 µL reactions to synthesize capped RNA.

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. The final concentration should be 25 µg/mL for PCR templates (compared to 50 µg/mL for plasmid templates).

A typical CleanScript IVT reaction following the kit protocol yields 0.8-1 mg of RNA per 100 µL of reaction using unmodified NTPs or N1-methylpseudouridine in place of UTP.

The standard protocol calls for a 3-hour incubation at 37°C, using either a thermal cycler with heated lid or a dry air incubator to prevent evaporation and condensation.

While DNase treatment is optional, it is recommended to remove template DNA. You can either use the one-pot DNase protocol in the product insert or purify the RNA and then follow a DNase treatment.

You can use lithium chloride precipitation, spin columns like QIAGEN RNeasy kits, or oligo(dT) if your RNA has a poly(A) tail. The choice often depends on your scale and downstream applications.

The control template is included to verify RNA transcription and aid with troubleshooting of technical issues that might arise. Following our recommended protocol, the template should yield 0.8-1 mg of 1.9 kb RNA transcript in 3 hrs of IVT. When using TriLink's FLuc control template, add 10 µL of the 0.5 µg/µL linearized plasmid provided for 5 µg total in a 100-µL IVT, as recommended in the protocol.

No, the FLuc plasmid template is provided linearized.

25 µg of linearized control template is provided. The amount is sufficient to use as a control for 5 x 100 µL reactions.

CleanCap AG (3' OMe) is a modified version of CleanCap AG to improve protein expression. CleanCap AG (3' OMe) includes the 5' N7-methyl-3'-O-methylguanosine commonly found in mRNA capped using anti-reverse cap analog (ARCA), whereas CleanCap AG has a naturally occurring 5' N7-methylguanosine structure.

The template must contain the correct T7 promoter sequence followed by an initiating sequence of 5' AGG 3' or 5' AGA 3' for proper use with CleanCap AG (3' Ome). Please refer to the protocol.

It is TriLink's proprietary in vitro transcription (IVT) method to produce high-quality, high-yield mRNAs from a broad range of sequences. It has been optimized to minimize dsRNA and improve in vivo protein expression from the resulting mRNAs. Please download the tech note to learn more.

Certificate of analysis

Intellectual property

support Contact us home Return to TriLink homepage