• Co-transcriptionally cap uridine-containing IVT RNA with Anti-Reverse Cap Analog (ARCA).
• Optimized for maximum transcript capping and yield.
• Produce up to 60 µg of ARCA-capped RNA from 1 µg of template in just 30 minutes.
• ScriptGuard™ RNase Inhibitor included to protect synthesized RNA from RNase degradation.

Product is for research use only (RUO)

Product Description:

The MessageMAX™ T7 ARCA-Capped Message Transcription Kit V2 is optimized for high-yield synthesis of canonical uridine-containing Anti-Reverse Cap Analog (ARCA)-capped RNA from an in vitro transcription (IVT) reaction. A 30 minute, 20 μl reaction yields up to 60 μg of ARCA-capped RNA from 1 μg of the control template DNA. ARCA ensures correct 5′ end incorporation, resulting in RNA that translates more efficiently in vivo than standard cap analogs. ScriptGuard™ RNase Inhibitor is included in the kit to protect synthesized RNA from RNase degradation.

The ARCA / NTP PreMix contains all four ribonucleotides (canonical nucleotides GTP, ATP, UTP and CTP) and the ARCA. The PreMix ensures the optimal concentration of each NTP and ratio of ARCA to GTP (4:1), maximizing transcript capping (~80%) and yield.

MessageMAX™ ARCA-capped RNA can be further processed into mRNA for expression in cells through the use of post-transcriptional capping using CELLSCRIPT™'s ScriptCap™ 2'-O-Methyltransferase (for conversion to a Cap 1 structure) and post-transcriptional tailing using A-Plus™ Poly(A) Polymerase Tailing Kit (available separately).

For reduction of immunogenicity, combine ARCA-capped mRNA with the Min-Immune™ Gold dsRNA Removal Kit to produce virtually dsRNA-free (to <0.005% of sample [LLOQ]), ultra-low immunogenicity mRNA that is suitable for downstream applications such as cell and gene therapy research and mRNA vaccine development.

CELLSCRIPT™ also offers the INCOGNITO™ T7 ARCA 5mC- & Ψ-RNA Transcription Kit to produce low immunogenicity ARCA-capped RNA containing the modified nucleotides 5-methyl-cytidine and pseudouridine.

Materials Supplied:

Important Store at -20°C in a freezer without a defrost cycle. Do not store at -70°C.

 

 

T7 Control Template DNA: Is a linearized 4.1 kb plasmid that contains a T7 promoter followed by a phage lambda dsDNA insert that encodes a 1,375 base runoff transcript. The Control Template DNA is provided at a concentration of 0.5 µg/µl in T₁₀E₁ Buffer (10 mM Tris-HCl, pH 7.5, 1 mM EDTA).

Materials Required, but not Supplied

• A DNA template for transcription of your RNA of interest
• Materials or kits for purification of the RNA product
• RNase-free TE Buffer (10 mM Tris-HCl, pH 7.5, 1 mM EDTA)
• Optional: TE saturated phenol/chloroform, 0.5-1 M EDTA

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Protocols for: MessageMAX™ T7 ARCA-Capped Message Transcription Kit 

 

Due to the constant updating of the protocols by the manufacturer we have provided a direct link to CELLSCRIPT’s product page, where the latest protocol is available.

Please note this will open a new page or window on your computer.

MessageMAX™ T7 ARCA-Capped Message Transcription Kit Protocol

Please note: all protocols are the responsibility of the product supplier

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

REFERENCES

1.Stepinski, J. et al., (2001) RNA 7, 1486.

2.Peng, Z-H et al., (2002) Org. Lett. 4, 161.

3.Jemielity, J. et al., (2003) RNA 9, 1108.

4.Warren, L. et al., (2010) Cell Stem Cell 7, 618.

5.Karikó, K. et al., (2008) Molecular Therapy 16, 1833.

6.Anderson, B.R. et al., (2010) Nucl. Acids Res. 17, 5884.

7.Karikó, K. et al., (2005) Immunity 23, 165.

8.Karikó, K. and Weissman, D. (2007) Curr. Opin. Drug Discov. Devel. 10, 523.

9.Sambrook, J. et al., (1989) Molecular Cloning: A Laboratory Manual (2nd ed.), New York, Cold Spring Harbor Laboratory Press.

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Anti-reverse cap analogs (ARCAs) are covered by U.S. Patent No. 7,074,596, and other patents issued or pending, licensed or assigned to CELLSCRIPT, Inc. These products are accompanied by a limited non-exclusive license for the purchaser to use the purchased product(s) solely for life science research. Purchase of these products does not grant rights to: (1) offer products, components of products or any derivatives thereof for resale; or (2) to distribute or transfer the products, components of products, or any derivatives thereof to third parties. Contact CELLSCRIPT for information on licenses for uses other than life science research

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

 

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200