- Synthesize up to 150 µg of uridine-containing IVT RNA (U-RNA) from 1 µg of template in two hours.
- Produce high yields of either long or short transcripts.
- Easily scale up reactions to produce milligram amounts of U-RNA.
- ScriptGuard™ RNase Inhibitor included to protect synthesized RNA from RNase degradation.
Product is for research use only (RUO)
Product Description:
The T7-Scribe™ Standard RNA IVT Kit is specially formulated to
produce high yields of canonical uridine-containing RNA (U-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 150 μg of
RNA from 1 μg of the control template and can be easily scaled up to
produce milligram amounts of U-RNA. These yields are made possible by
the high-performance properties of the T7-Scribe™ enzyme. The T7-Scribe™
Standard RNA IVT Kit produces high yields of either long or short
transcripts and can be readily modified to prepare fluorescent-,
biotinylated- or digoxigenin-labeled RNA. ScriptGuard™ RNase Inhibitor is included in the kit to protect synthesized RNA from RNase degradation.
T7-Scribe™ IVT U-RNA can be further processed into U-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 reduction of immunogenicity, combine U-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™ line of products
to produce low immunogenicity mRNA containing the modified nucleotides
N1-methyl-pseudouridine, pseudouridine and/or 5-methyl-cytidine.
Product Performance:
The standard two hour, 20 μl reaction will yield up to 150 μg of RNA
from 1 μg of the control template. However, lower amounts of DNA
template can be used successfully in a T7-Scribe™ reaction. Table 1
summarizes the yields obtained when reducing the amount of control
template DNA in a T7-Scribe™ reaction. Results may vary depending on the
template used. Increasing the reaction time to four to six hours may
increase the yield of RNA.
Table 1. RNA yields from T7-Scribe™ Standard RNA IVT Kit reactions using different DNA inputs. The standard protocol uses 1 µg of DNA input.
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)
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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