| Field | Specification |
|---|---|
| Mfr No | |
| Product Type | |
| Shipping | |
| Source | Recombinant (E. coli) |
| Storage |
This is an engineered T7 RNA polymerase variant (low dsRNA) derived from the wild-type T7 RNA polymerase and produced in Escherichia coli. It significantly reduces the production of double-stranded RNA (dsRNA) while efficiently incorporating cap analogs, and exhibiting highly efficient in vitro transcription (IVT) comparable to the wild-type T7 RNA polymerase. It catalyzes the 5'→3' synthesis of RNA on double-stranded DNA from its T7 promoter sequence (5'-TAATACGACTCACTATAG*-3') and uses NTPs as substrates.
Note: G* is the first base of the RNA transcript.
Feature
- dsRNA level lower to about 1/100000
- Comptiable with Trilink CleanCap AG, LZCap
- High yields comparable to WT
- Lower Cap input
- Animal origin-free (AOF)
Please find information on the development of this enzyme.
Components
|
Components No. |
Name |
10629ES10 |
10629ES60 |
10629ES72 |
10629ES86 |
|
|
|
(10 KU) |
(100 KU) |
(250 KU) |
(2,500 KU) |
|
10629 |
Hieff™ T7 RNA Polymerase (GMP Grrade, low dsRNA, 250 U/μL) |
40 μL |
400 μL |
1 mL |
10 mL |
Specifications
|
Source |
Recombinant E. coli with T7 RNA Polymerase gene |
|
Optimum Temperature |
37℃ |
|
Storage Buffer |
50 mM Tris-HCl, 1 mM EDTA, 10 mM DTT, 100 mM NaCl, 0.1% Triton X-100，50% (v/v) glycerin，pH7.9 at 25℃ |
|
Unit Definition |
The amount of enzyme required to incorporate 1 nmol of [3H] GMP into the acid-insoluble precipitate within 1 hour at 37°C and pH 8.0 is defined as 1 unit. |
|
Recommended Mg2+ |
30mM Magnesium Acetate |
Glycerol Content |
Contains Glycerol |
QC Standard
|
Items |
Specification/Standard |
|
Enzyme activity |
250-300U/μL |
|
Protein Purity |
≥95% |
|
Endotoxin |
<20 EU/mg |
|
Protease |
Negative |
|
Exonuclease |
Negative |
|
Nickase |
Negative |
|
RNase |
Negative |
|
E.coli host protein |
<50ppm |
|
E.coli host DNA |
<10 fg/U |
|
Mycoplasma examination |
Negative |
Related Products
Tris NTPs synergistically decreasing dsRNA
Figures
Figure 2. Evaluation of the immunogenicity of the IVT products in murine RAW264.7 cells (Figures 2A and 2B). IFN-β mRNA and protein levels were reduced in RAW264.7 cells transfected with mRNA produced by mutants compared to wild-type, indicating that mRNA synthesized by the wild-type T7 RNAP elicited the strongest immune response, while mRNA from the mutants showed a significantly reduced response.
Figure 3. The dsRNA content in mRNA synthesized with CleaScript™ T7 RNA polymerase is lower than that in mRNA synthesized with the wild-type enzyme after cellulose treatment.
Shipping and Storage
The products are shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.
Publication:
Engineered T7 RNA polymerase reduces dsRNA formation by lowering terminal transferase and RNA-dependent RNA polymerase activities, The FEBS Journal 03 March, 2025
Store this enzyme at –20°C and avoid repeated freeze-thaw cycles to preserve catalytic activity. The product is shipped on dry ice and remains stable for up to one year from the date of manufacture when stored under recommended conditions. Aliquoting the stock solution into single-use volumes is recommended for enzymes used infrequently to minimize thermal cycling of the bulk stock.
This enzyme is validated for use in molecular biology research applications. It is suitable for use in both standard laboratory research settings and, where applicable, optimized reaction workflows requiring high specificity, sensitivity, or throughput.
One unit (U) is defined as the amount of enzyme that catalyzes the conversion of a defined quantity of substrate under standardized conditions of temperature, pH, ionic strength, and substrate concentration. Refer to the product Certificate of Analysis for lot-specific activity data.
This enzyme is produced as Recombinant (E. coli) and supplied as a Research Use Only (RUO) reagent. Each lot is subjected to activity assay, purity assessment by SDS-PAGE, and functional validation prior to release. A Certificate of Analysis (CoA) and Safety Data Sheet (SDS) are available on request.
This enzyme is produced recombinantly in E. coli under stringent quality controls and validated on multiple substrate types. Key performance parameters include specific activity, substrate specificity, cofactor requirements, optimal pH/temperature range, and compatibility with downstream assays. Refer to the product datasheet for validated lot-specific activity data and recommended reaction conditions.
Yeasen Biotechnology supports custom enzyme solutions across multiple service lines — from GMP-grade bulk supply to directed enzyme engineering. Contact BioHippo to discuss requirements and initiate a project inquiry.
▶ GMP-Grade & Bulk Supply
Select Yeasen enzymes are available in GMP grade, manufactured in an ISO 13485-certified UCF.ME™ ultra-clean molecular enzyme facility with FDA Drug Master File (DMF) support.
- GMP-grade release testing and CoA documentation
- ISO 13485-certified production facility
- Scalable from milligram to multi-gram quantities
- Consistent lot-to-lot activity specifications
▶ Glycerol-Free & Custom Formulation
Glycerol-free enzyme formats are available for applications requiring lyophilization compatibility, liquid handling automation, or direct IVD master mix integration.
- Glycerol-free liquid format (standard and custom buffers)
- Lyophilization-ready enzyme preparation
- Custom reaction buffer optimization for specific assay conditions
- Compatible with freeze-drying workflows for point-of-care formats
▶ Molecular IVD RDC Service
Yeasen's Research and Development Contracting (RDC) team delivers end-to-end solutions for molecular diagnostic product development, covering enzyme selection through clinical validation support.
- Enzyme selection and performance matching
- Primer/probe design and reaction buffer optimization
- Sensitivity, specificity, and precision validation studies
- Stability studies and SNP evaluation
- Instrument platform compatibility assessment
▶ ZymeEditor™ Enzyme Engineering
Yeasen's proprietary ZymeEditor™ directed evolution and rational design platform enables the development of custom enzyme variants with tailored performance characteristics not available in off-the-shelf products.
- Directed evolution for enhanced thermostability, processivity, or fidelity
- Rational design for altered substrate specificity or cofactor requirements
- Library screening from Yeasen's proprietary enzyme variant collection
- Scale-up to commercial quantities upon candidate confirmation
ⓘ Customization services are fulfilled by Yeasen Biotechnology. Lead times and minimum order quantities vary by service type. Contact BioHippo for project scoping and pricing.
