The INCOGNITO™ SP6 Ψ-RNA Transcription Kit is optimized for high-yield synthesis of pseudouridine-containing RNA (Ψ-RNA) from an in vitro transcription (IVT) reaction. Although yield varies with the DNA template and other factors, a standard two hour, 20 μl reaction will yield up to 60 μg of RNA from 1 μg of the control template thanks to the high-performance properties of the SP6-Scribe™ enzyme. The standard reaction can be scaled up to produce milligram amounts of RNA containing the canonical nucleotides ATP, CTP, GTP and the modified nucleotide pseudouridine-5'-triphosphate (ΨTP).

Modified Ψ-mRNAs translate into higher protein levels and lower innate immune responses in human and other mammalian cells that express various RNA sensors compared to unmodified mRNAs.

INCOGNITO™ SP6 IVT Ψ-RNA can be further processed into low immunogenicity mRNA (5'-end capped and 3'-end poly[A] tailed) through the use of CELLSCRIPT™'s ScriptCap™ Cap 1 Capping System (contains both ScriptCap™ Capping Enzyme and 2'-O-Methyltransferase) and A-Plus™ Poly(A) Polymerase Tailing Kit (available separately).

For maximum reduction of immunogenicity, combine INCOGNITO™ Ψ-mRNA with the Min-Immune™ Gold dsRNA Removal Kit to produce virtually dsRNA-free, ultra-low immunogenicity mRNA that is suitable for downstream applications such as cell and gene therapy and mRNA vaccines.

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

Materials Supplied:

SP6 Control Template DNA: Is a linearized 4.1 kb plasmid including an SP6 promoter followed by a 1.4 kb lambda DNA insert, resulting in a 1,386 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

Terms and Conditions: The Product listed is currently offered by CELLSCRIPT™ for research use only under the defined Terms and Conditions.

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• Manual
• SDS

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Important

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

Uses and Label Licenses for Specific Products

INCOGNITO™ kits or other products or services comprising N1-methyl-pseudouridine, pseudouridine, 5-methyl-cytidine and/or other modified nucleotides for use in making RNA or mRNA (the “Products”) that are covered by technologies and IP disclosed in U.S. Patent Nos. 8,278,036 and 9,750,824 and PCT Publ. Nos. WO 2007/024708 and WO/2011/071931 and U.S. and international divisional, continuation, or other patents and patent applications derived therefrom that are owned by the University of Pennsylvania and licensed to CELLSCRIPT™ (“Products”) are sold for research use only.

By purchasing these Products from CELLSCRIPT™ or an authorized distributor of CELLSCRIPT™, Purchaser receives a limited non-exclusive, non-transferable right to use the Products purchased from CELLSCRIPT™ solely for its own internal laboratory research use (the “Product Use”). Such Product Use expressly excludes any Commercial Use comprising [A] clinical or other use in humans, [B] veterinary, livestock, agricultural or commercial use in animals, and/or [C] manufacture, distribution, importation, exportation, or sale of any products and/or services made using Products, it being understood that [A]–[C] are separately and/or collectively defined as a “Commercial Use” herein. By purchase of Products, Purchaser agrees not to make, import, use, transfer, offer for sale, or sell Products, components of Products, or derivatives of Products, including modified nucleoside-containing RNA or cells made by use of Products, or to provide services, information, or data obtained by the use thereof in exchange for money or other consideration. CELLSCRIPT™ provides no warranties (statutory or implied) concerning non-infringement of intellectual property rights of any other parties, and all such warranties are expressly disclaimed. Please contact CELLSCRIPT™ to inquire about a license for a Commercial Use.

The license remains in effect as long as the product is being used in accordance with the license terms. The license is terminated if the user breaches any of the license terms, such as using the product for commercial purposes without proper authorization.

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• Optimized for high-yield synthesis of pseudouridine-containing IVT RNA (Ψ-RNA).
• Produce up to 60 µg of Ψ-RNA from 1 µg of template in two hours with an SP6 promoter.
• Easily scale up reactions to produce milligram amounts of Ψ-RNA.
• Lower immunogenicity in mammalian cells through incorporation of Ψ in synthesized mRNA.

Product is for research use only (RUO)

Use and Label Licenses for Specific Products:

Purchaser receives a limited non-exclusive, non-transferable right to use the Products purchased from CELLSCRIPT™ solely for its own internal laboratory research use. See the Label License tab for more information.

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