- Optimized for high-yield synthesis of pseudouridine-containing IVT RNA (Ψ-RNA).
- Produce up to 100 µg of Ψ-RNA from 1 µg of template in two hours.
- 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.
Product Description:
The INCOGNITO™ T7 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 100 μg of
RNA from 1 μg of the control template thanks to the high-performance
properties of the T7-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™ T7 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
research and mRNA vaccine development.
Important Store at -20°C in a freezer without a defrost cycle. Do not store at -70°C.
| INCOGNITO™ T7 Ψ-RNA Transcription Kit Contents (10 reactions) |
|
Kit Component |
Reagent Volume |
|
T7-Scribe™ Enzyme Solution |
20 μl |
|
10X T7-Scribe™ Transcription Buffer II |
20 μl |
GAΨC PreMix (25 mM each GTP, ATP, ΨTP & CTP) |
60 μl |
|
100 mM Dithiothreitol (DTT) |
20 μl |
|
RNase-Free DNase I, 1 U/μl |
10 μl |
|
ScriptGuard™ RNase Inhibitor, 40 U/μl |
10 μl |
|
T7 Control Template DNA, 0.5 μg/μl |
10 μl |
|
RNase-Free Water |
1.4 ml |
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)
Terms and Conditions: The Product listed is currently offered by CELLSCRIPT™ for research use only under the defined Terms and Conditions.
If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200