Product Number: TR03

Shipping and Storage

Store at -20°C℃±5℃.

Component

Description

As a biomacromolecule, mRNA can be synthesized at scale through in vitro transcription (IVT). The T7 promoter is currently one of the most efficient promoters for transcription, enabling rapid and straightforward production of large quantities of RNA molecules using T7 RNA polymerase. As a byproduct of IVT, double-stranded RNA (dsRNA) can be recognized by corresponding nucleic acid receptors, triggering innate immune inflammatory responses that severely impact the efficacy of mRNA vaccines. Strict control during production is essential. This product employs a T7 RNA polymerase engineered through molecular evolution, significantly reducing dsRNA levels in transcription products and lowering the immunogenicity of synthesized mRNA.

This product is a GMP-grade recombinant T7 RNA polymerase produced through large-scale fermentation using Escherichia coli. It is manufactured with pharmaceutical-grade raw materials and excipients, with strict control over host protein residues, nucleic acid residues, and other impurities. The production and quality management processes comply with GMP standards, ensuring traceability of the entire production process and all raw materials.

Application

1. Synthesize single stranded RNA for the preparation of mRNA vaccines and other applications.
2. Synthesize highly specific RNA probes. 
3. Synthesize siRNA precursor.
4. Preparation of RNA splicing precursors.
5. Synthesize capped RNA using cap analogues.

Quality control

Storage buffer solution

50mM Trizma base; 100mM NaCl; 1mM EDTA; 20mM β-ME; 50% (v/v) Glycerol; 0.1% Triton X-100; pH 7.5.

Source

E.coli carrying bacteriophage T7 RNA polymerase gene

Features

It exhibits high specificity for the T7 promoter.

Definition of active units

The enzyme amount required to incorporate 1nmol of [³H] GMP into an acid insoluble precipitate within 1 hour at 37℃ and pH 8.0 is defined as 1 active unit.

Related Products

TR01 – T7 RNA Polymerase

TR02 – Thermostable T7 RNA Polymerase

TR03 – T7 RNA Polymerase 200U/ul

GMP-T701 – T7 RNA Polymerase, GMP Grade

E131 – T7 High Yield RNA Transcription kit

Product Number: GMP-T701            Animal-free          Ampicillin-free

Shipping and Storage

At -20±5℃.

Description

As a biological macromolecule, mRNA can be synthesized on a large scale by in vitro transcription (IVT). T7 promoter is one of the most efficient promoters at present. Therefore, T7 RNA polymerase can be used for in vitro transcription to obtain more synthetic products. T7 RNA polymerase is a T7 promoter-specific, DNA-dependent, 5’→3′ RNA polymerase from T7 bacteriophage. Usingdouble stranded DNA as the template, it transcribes RNA complementary to the single stranded DNA located at the downstream of T7promoter. T7 RNA polymerase has been commonly used for in vitro mRNA synthesis.

The polymerase is GMP Grade produced in E. coli. Our manufacturing processes are strictly controlled to ensure the end products free from host protein or nucleic acid contaminations and other impurities following the Pharmaceutical Manufacturing Guidelines. We guarantee the manufacturing and quality control comply with GMP regulation for tracking each and every step of the manufacturing process, including raw material sourcing.

This product has completed the DMF record of FDA and passed the HALAL certification.

Quality Elements

Annotation: ChP refers to the Pharmacopoeia of the People’s Republic of China.

Complying to following regulations

1. ISO 9001:2015, certified facility.
2. 《GMP Appendix – Cellular therapeutic product》National Medical Products Administration.
3. 《The Pandect of Genetic Therapeutic Product for Human》Chinese Pharmacopoeia Commission.
4. USP Chapter <1043>, Ancillary Materials for Cell, Gene, and Tissue-Engineered Products.
5. USP Chapter <92>, Growth Factors and Cytokines Used in Cell Therapy Manufacturing.
6. Ph. Eur. General Chapter 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products.

Feature

Highly specific for T7 promoter, suitable for RNA in vitro synthesis.

Application

1. Single stranded RNA synthesis
2. RNA probe synthesis.
3. siRNA precursor synthesis
4. Precursor for RNA splicing preparation
5. Capped RNA synthesis.

Examples

Fig: RNA transcription in vitro.

1. From left to right, the quantity of T7 RNA Polymerase copies were 20U, 4U, 0.8U.
2. DNA templates were segments of 2Kb.

Unit definition

At 37℃, pH8.0, within 1 hour, the amount of enzyme required that will incorporate 1nmol tritium labeled GMP into acid-insoluble material is defined as one unit of enzyme activity.

Storage buffer

100mM NaCl; 50mM Tris-HCl (pH 7.9); 1mM EDTA; 20mM 2-mercaptoethanol; 0.1% Triton X-100; 50% (v/v) Glycerol。

Related Products

TR01 – T7 RNA Polymerase

TR02 – Thermostable T7 RNA Polymerase

TR03 – T7 RNA Polymerase 200U/ul

GMP-T701 – T7 RNA Polymerase, GMP Grade

E131 – T7 High Yield RNA Transcription kit

Product Number: TR01

Shipping and Storage

-20℃.

Components

Description

T7 RNA polymerase is a DNA dependent 5’→ 3’rna polymerase that highly specifically recognizes the T7 promoter sequence. T7 RNA polymerase can catalyze the incorporation of NTP downstream of T7 promoter of single stranded or double stranded DNA to synthesize RNA complementary to template DNA downstream of T7 promoter.

Source

It is expressed by Escherichia coli and the expressed gene is bacteriophage T7 RNA polymerase gene.

Concentration

50U/μl

Purity

No DNA endonuclease and exonuclease, no RNase.

Feature

T7 RNA polymerase can recognize modified NTPs, such as biotin labeled, digoxigenin labeled, fluorescein labeled NTPs, and can be used for the synthesis of various labeled RNAs. At the same time, it has high specificity for T7 promoter.

Application

For RNA synthesis, the synthesized RNA can be used or used as: hybridization probe, genomic DNA sequence analysis, RNase protection assay, antisense RNA synthesis, as RNA template for in vitro translation, substrate for RNA splicing research, RNA secondary structure and RNA protein interaction, nucleic acid amplification analysis, siRNA, miRNA and other small RNAs.

Unit definition

The amount of enzyme required to catalyze the incorporation of 1 nmol of AMP into polynucleotides within 60 min at 37 ° C was defined as 1 active unit.

Inactivation or inhibition

Heating at 70℃ for 10 min can inactivate T7 RNA polymerase. Addition of an appropriate amount of EDTA can also inactivate T7 RNA polymerase. Chelating agents, sodium, potassium or ammonium salts with concentrations greater than 150mm can significantly inhibit the activity of T7 RNA polymerase.

Related Products

TR01 – T7 RNA Polymerase

TR02 – Thermostable T7 RNA Polymerase

TR03 – T7 RNA Polymerase 200U/ul

GMP-T701 – T7 RNA Polymerase, GMP Grade

E131 – T7 High Yield RNA Transcription kit

Product Number: E131

Shipping and Storage

-20℃

Component

Description

As biological macromolecules, mRNA can be synthesized on a large scale through in vitro transcription (IVT,in vitro transcription). The T7 promoter is currently one of the most efficient promoters for transcription, so using T7 RNA polymerase (TR01,T7 RNA Polymerase) for in vitro transcription can easily and quickly obtain a large number of RNA molecules.This kit has been optimized through a series of transcription reaction systems, and RNA complementary to a single strand in DNA is synthesized from downstream of the template DNA T7 promoter using T7 RNA Polymerase. The operation is simple and fast.

This reagent kit can transcribe and produce 150-200μg RNA in one reaction. The synthesized RNA can be used for downstream applications such as RNA structure and function research, RNA enzyme protection, probe hybridization, RNAi, microinjection, and in vitro translation.

Application

1. Synthesis of single stranded RNA;
2. Synthesis of highly specific RNA probes;
3. Synthesis of siRNA precursors;
4. Produce precursors for RNA splicing reactions.

Related Products

TR01 – T7 RNA Polymerase

TR02 – Thermostable T7 RNA Polymerase

TR03 – T7 RNA Polymerase 200U/ul

GMP-T701 – T7 RNA Polymerase, GMP Grade

E131 – T7 High Yield RNA Transcription kit

Product Number: TR02

Shipping and Storage  

-20℃

Components

Description

Thermalstable T7 RNA Polymerase is a genetically modified T7 RNA polymerase.This enzyme highly specifically recognizes the T7 promoter sequence and can perform efficient in vitro transcription at 50℃.During the in vitro transcription process, it can improve the capping efficiency and eliminate the production of dsRNA byproducts.

Features

The T7 promoter has high specificity and is used for the synthesis of in vitro RNA (including small RNAs).

Application

1. Synthesis of single stranded RNA;
2. Synthesis of highly specific RNA probes;
3. Synthesis of siRNA precursors;
4. Produce precursors for RNA splicing reactions;
5. Using cap analog as a primer, produce Capped mRNA.

Unit definition

The amount of enzyme required to mix 1 nmol of [³H] GMP with acid insoluble precipitate within 1 hour at 50℃ and pH 8.0 is defined as 1 active unit.

Quality Control

After multiple column purification, SDS-PAGE gel detection only showed clear and single target bands, with a purity of 95%. PCR detection showed no residual Escherichia coli DNA and no contamination of RNase, nucleic acid endonucleases, and exonucleases.

Suggestions

1. For effective transcription in specific regions, it is recommended to pre cut the template DNA into flat or 5 ‘protruding ends downstream of the region.
2. The binding of spermidine in the buffer with nucleic acid may form insoluble substances, and it is recommended to add template DNA finally.

Related Products

TR01 – T7 RNA Polymerase

TR02 – Thermostable T7 RNA Polymerase

TR03 – T7 RNA Polymerase 200U/ul

GMP-T701 – T7 RNA Polymerase, GMP Grade

E131 – T7 High Yield RNA Transcription kit