Product Overview
Hi-yield T7 in vitro transcription reagent (low dsRNA) is designed for in vitro transcription of RNA with low dsRNA byproduct using T7 RNA polymerase mutant with optimized transcription reaction system.
The kit is capable of synthesizing mRNA using linear double-stranded DNA containing T7 promoter as template and NTPs as substrate.
The kit is suitable for incorporation of modified nucleotides to obtain biotin labeled, dye labeled and radiolabeled RNA. The kit also can be applied in synthesis of co-transcriptionally capping mRNAs with cap analogs. The kit contains modified nucleotides N1-Me-pUTP as alternative.
Each standard reaction yields up to 150-200 μg of RNA from 1μg DNA template. Reaction size can be scaled up to yield milligram-level RNA as needed.
RNA synthesized from the kit is suitable for many applications including RNA structure and function studies, ribozyme biochemistry, RNase protection assays and hybridization-based blots, anti-sense RNA and RNAi experiment.
RNA synthesized from the kit also is suitable for enzymatic production of capped RNA by vaccinia capping enzyme (HBP000606) and 2’-O-Methyltransferase (HBP000701) and being tailed up with Poly(A) polymerase (HBP000801) to improve the stability and translational competence of RNA used for in vitro translation, transfection, and microinjection.
Key Features
High Yield RNA Synthesis
Delivers exceptional transcription efficiency, generating 180-220 μg of RNA from 1 μg of DNA template under standard reaction conditions, enabling robust protein expression and scalable RNA production.
Low dsRNA Byproduct Formation
Powered by the engineered Low dsRNA T7 RNA Polymerase, the kit produces more than 10-fold less dsRNA than wild-type T7 RNAP, facilitating downstream purification and supporting the development of high-quality mRNA products.
Broad Compatibility with Modified IVT Workflows
Compatible with a wide range of modified nucleotides, cap analogs, and co-transcriptional capping strategies, enabling the synthesis of diverse RNA modalities tailored to specific application needs.
Flexible Template Compatibility
Efficiently transcribes DNA templates ranging from 200 bp to 12 kb, supporting applications from conventional mRNA production to long and structurally complex RNA constructs.
Ready-to-Use, Pre-Optimized System
Supplied as a complete IVT solution with all essential reagents included, eliminating manual reagent preparation and reaction optimization to streamline workflow setup and improve reproducibility.
Applications
GMP Manufacturing and Preclinical Development
Suitable for the development and production of mRNA vaccines, gene therapies, and other RNA therapeutics in both preclinical and GMP manufacturing settings.
Reduced Purification Burden for Low-dsRNA mRNA Production
The kit incorporates the engineered Low dsRNA T7 RNA Polymerase (Cat. No. HBP000340), generating substantially lower dsRNA levels than conventional T7 systems. Compared with the T7 RNA Polymerase mutant (Cat. No. HBP000330), dsRNA content is reduced by more than 50%, helping simplify downstream purification and reduce manufacturing costs.
High-Dose mRNA Therapeutic Development
By minimizing dsRNA impurities at the source, the kit supports the generation of mRNA products with improved safety profiles, enabling the exploration of higher dosing strategies in therapeutic applications.
Accelerated IVT Workflow Development
As a complete, pre-optimized transcription system, the kit significantly reduces workflow setup time and eliminates the need for extensive condition optimization, allowing users to rapidly obtain high-quality IVT products with excellent reproducibility.
Performance Data
Reduces dsRNA content at 37℃
Validated against multiple templates, the M13 mutant has a significant advantage in reducing dsRNA content.
Lower immunogenicity
Template 1 and template 2 with co-transcription capping using M13 resulted in lower double-stranded content and transfection into RAW cells produced lower levels of the IFN-α and IL-6.
Component List
| Component | Concentration | Volume |
| ATP | 100 mM | 100 μL |
| CTP | 100 mM | 100 μL |
| GTP | 100 mM | 100 μL |
| UTP | 100 mM | 100 μL |
| N1-Me-pUTP | 100 mM | 100 μL |
| 10×IVT Buffer 9 | / | 100 μL |
| Enzyme Mix (low dsRNA) | / | 100 μL |
| Control template 1 | 0.5 μg/μL | 10 μL |
| LiCl | 5 M | 1.4 mL |
| DNase I | 5 U/μL | 100 μL |
FAQ
How does this kit reduce dsRNA byproducts?
The kit incorporates an engineered low dsRNA T7 RNA Polymerase, generating significantly fewer dsRNA impurities than conventional T7 IVT systems and reducing downstream purification burden.
What RNA yield can be achieved with this kit?
Under standard reaction conditions, the kit can generate up to 180-220 μg of RNA from 1 μg of DNA template, and reactions can be scaled up to produce milligram quantities of RNA.
Is the kit compatible with modified nucleotides and cap analogs?
Yes. The kit supports a broad range of modified nucleotides and cap analogs for the synthesis of labeled and co-transcriptionally capped RNAs.
What applications is this kit suitable for?
The kit is suitable for mRNA vaccine and gene therapy development, as well as other RNA applications requiring high-yield, low-dsRNA IVT products.
How should the kit be stored and transported?
The kit should be transported below 0°C and stored at -25 to -15°C.
