New England Biolabs Canada
Expand menu
Sign In or Sign UpMy NEB

HiScribe® T7 mRNA Kit with CleanCap® Reagent AG ...

Catalog # Concentration Size List Price Quantity Your Price
E2080L 100 reactions $3,526.00
$3,173.40
E2080S 20 reactions $882.00
$793.80
Catalog # Size List Price Your Price
E2080L 100 reactions $3,526.00
$3,173.40
E2080S 20 reactions $882.00
$793.80
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
  • Generate up to 90 µg of Cap-1 mRNA per reaction from 1 µg of control template
  • NTPs are provided separately to enable partial or full substitution of modified NTPs including N1-Methyl-Pseudouridine-5’-Triphosphate (NEB #N0431), 5-Methyl-Cytidine-5’-Triphosphate (NEB #N0432), Pseudouridine-5’-Triphosphate (NEB #N0433) and 5-Methoxy-Uridine-5’-Triphosphate (NEB #N0434)
  • Template removal and RNA purification reagents included
  • Includes linearized control template for use with CleanCap® Reagent AG for verification of RNA synthesis
  • Getting ready to scale up RNA synthesis? Download our new technical note “Scaling of High-Yield In vitro Transcription Reactions for Linear Increase of RNA Production” for a generalized set of recommendations for synthesizing high yields of RNA.

Most eukaryotic mRNAs require a 7-methyl guanosine (m7G) cap structure at the 5´-end and a poly(A) tail at the 3′-end for efficient translation.  The HiScribe T7 mRNA Kit with CleanCap Reagent AG utilizes an optimized RNA synthesis formulation and trinucleotide cap analog technology for co-transcriptionally capping mRNAs that contain a natural Cap-1 structure in a single simplified reaction without compromising RNA yield.  By using a DNA template with a T7 promoter sequence followed by an AG initiation sequence and an encoded poly(A) tail, mRNAs can be transcribed with a 5´-m7G Cap-1 structure that is polyadenylated, translationally competent and able to evade the cellular innate immune response.  

The HiScribe T7 mRNA Kit with CleanCap Reagent AG is formatted with individual vials of NTPs and CleanCap Reagent AG to allow for partial or complete substitution of modified NTPs, with a total kit yield of 1.8 mg of mRNA.  Cap-1 mRNA synthesized from this kit is suitable for many applications, including transfections, microinjections, in vitro translation, preclinical mRNA therapeutic mRNA studies as well as RNA structure and function analysis.

This kit contains sufficient reagents for 20 reactions (20 µl each).  Each standard reaction yields ≥ 90 µg of RNA from 1 µg CLuc AG Control Template DNA.  Each kit can yield ≥ 1.8 mg RNA.

Figure: 1: CleanCap Reagent AG





FIgure 2: Schematic of CleanCap Reagent AG Promoter and Initiation Sequence





Figure 3: CleanCap Reagent AG results in higher mRNA synthesis yield than the ARCA analog

 

All reactions were performed with 5 mM CTP, 5 mM UTP and 6 mM ATP.  Standard IVT reactions included 5 mM GTP and no cap analog.  ARCA reactions contained a 4:1 ratio of ARCA:GTP (4mM:1mM).  IVT with CleanCap Reagent AG contained 5 mM GTP and 4 mM CleanCap Reagent AG and was performed according to recommended protocol (Standard mRNA Synthesis, HiScribe T7 mRNA Kit with CleanCap Reagent AG).  Reactions were incubated for 2 hours at 37°C, purified and quantified by NanoDrop®.


 
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  T7 RNA Polymerase Mix M0255AAVIAL -20 1 x 0.04 ml
  LiCl Solution B2051AVIAL -20 1 x 1.4 ml
  DNase I (RNase-free) M0303AAVIAL -20 1 x 0.04 ml 2,000 units/ml
  CLuc AG Control Template N2078AVIAL -20 1 x 0.02 ml 0.25 mg/ml
  10X T7 CleanCap® Reagent AG Reaction Buffer B2082AVIAL -20 1 x 0.04 ml
  CleanCap® Reagent AG S1413AVIAL -20 1 x 0.04 ml 40 mM
  ATP N0489AVIAL -20 1 x 0.04 ml 60 mM
  GTP N0490AVIAL -20 1 x 0.04 ml 50 mM
  CTP N0491AVIAL -20 1 x 0.04 ml 50 mM
  UTP N0492AVIAL -20 1 x 0.04 ml 50 mM
  Dithiothreitol (DTT) B1222AVIAL -20 1 x 0.5 ml 100 mM
  T7 RNA Polymerase Mix M0255AVIAL -20 2 x 0.1 ml
  LiCl Solution B2051AVIAL -20 2 x 1.4 ml
  CLuc AG Control Template N2078AVIAL -20 1 x 0.02 ml 0.25 mg/ml
  DNase I (RNase-free) M0303AVIAL -20 2 x 0.1 ml 2,000 units/ml
  Dithiothreitol (DTT) B1222AVIAL -20 1 x 0.5 ml 100 mM
  10X T7 CleanCap® Reagent AG Reaction Buffer B2082AAVIAL -20 1 x 0.2 ml
  ATP N0489AAVIAL -20 1 x 0.2 ml 60 mM
  GTP N0490AAVIAL -20 1 x 0.2 ml 50 mM
  CTP N0491AAVIAL -20 1 x 0.2 ml 50 mM
  UTP N0492AAVIAL -20 1 x 0.2 ml 50 mM
  CleanCap® Reagent AG S1413AAVIAL -20 1 x 0.2 ml 40 mM
Features
  • Streamlined workflow with single-step co-transcriptional capping
  • CleanCap® Reagent AG trinucleotide cap technology results in a natural Cap-1 structure, maximizing translatability and minimizing immune response from synthetic mRNA
  • High capping efficiency (> 95% capped material1)
  • Optimized for high yields
  • Suitable for full- or partial- modified nucleotide substitution

1Final capping is dependent upon the CleanCap Reagent, DNA template and final mRNA sequence. Secondary structure due to RNA length and base composition can affect final capping efficiency. 

 
An exception occurred during the operation, making the result invalid. Check InnerException for exception details.

Companion Products
Quality Control Assay
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
Certificate of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
Legal And Disclaimer

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.CleanCap® and products incorporating it are sold under license from TriLink BioTechnologies, LLC and may be used for research use only, not for use in diagnostics, therapeutics procedures or for use in humans. For additional information, please see the product’s Limited Use License at https://www.trilinkbiotech.com/cleancap-research-license

Other Products You May Be Interested In
 Top