Description

S-adenosylmethionine is the most important methyl donor in vivo and an auxiliary substrate involved in methyl transfer reaction. In the synthesis of mRNA in vitro,both the hydroxyl group of ribose and the amino group of base need the participation of SAM during methylation.

Feature

The SAM is prepared in 0.005M H₂SO₄ and 10% EtOH solution, filtered to provide in the form of aseptic liquid. In the process of mRNA capping, the combination of SAM and methyltransferase can transform the Cap0 structure of the 5′ cap region into Cap1 structure.

Specification

Customized Spesfication:

1）Endonuclease Activity (Nicking) – A 50 µl reaction in Buffer 2 containing 1 µg of supercoiled PhiX174 DNA and a minimum of 5 µl of S-adenosylmethionine (SAM) incubated for 4 hours at 37ºC results in <20%conversion to the nicked form as determined by agarose gel electrophoresis.

2）Non-Specific DNase Activity (16 Hour) – A 50 µl reaction in NEBuffer 2 containing 1 µg of PhiX174-HaeIII DNA and a minimum of 5 µl of S-adenosylmethionine (SAM) incubated for 16 hours at 37ºC results in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.

3）Restriction Digest (CpG Resistant, SAM) – A 20 µl reaction in 1X Buffer 2 containing 1 µg of LambdaDNA, 1 unit of M. SssI (CpG Methyltransferase), and 160 µM S-adenosylmethionine (SAM) is incubated for 1hour at 37°C. The resulting DNA is resistant to digestion with BstUI as determined by agarose gel electrophoresis

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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 Description

Vaccinia Capping enzyme is a recombinant protein from Vaccinia virus. Vaccinia Capping Enzyme can add 7-methylguanylate cap structure (Cap 0) to the 5′ end of RNA. In eukaryotes, these terminal cap structures are involved in stabilization, transport, translation of mRNAs. Vaccinia Capping enzyme is composed of two subunits (D1R, D12L), and has three enzymatic activities (RNA triphosphatase, guanylyltransferase by the D1R subunit and guanine methyltransferase by the D12L subunit). All necessary for addition of a complete Cap 0 structure, m7Gppp5’N). In vitro, transcripts can be capped in the presence of reaction buffer, GTP, and the methyl donor (SAM).

Source

Recombinant E.coli

Concentration and Size

10U/μL

Unit Definition

One unit is defined as the amount of enzyme required to incorporate 10pmol of GTP into an 80 nt transcript in 1 hour at 37℃.

Storage Buffer

20mM Tris-HCl (pH 8.0), 0.1mM EDTA, 100mM NaCl, 1mM DTT, 0.1%(v/v) Triton X-100, 50%(v/v) glycerol.

Storage Conditions

-20 °C

Quality Assurance

Free of Endonuclease, Exonuclease, and RNase activities.

Physical Purity

Purity >95% by SDS-PAGE.

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Description

Restriction endonucleases are a class of endonuclease that recognize specific deoxynucleotide sequences and cleave the phosphodiester bond between two deoxyribonucleotides at specific sites in each chain. BsaI derived from Bacillus Thermophilus, is a commonly used restriction endonuclease. BsaI recognizes the following sequence:

5’…GGTCTC(N)1↓…3’;

3’…CCAGAG(N)5↑…5’.

Then it functions DNA cleavage.

This product uses recombinant protein production technology to obtain restriction endonuclease BsaI. Applying pharmaceutical leveled adjuvant and material for production, strictly controlling host protein residues, nucleic acid residues and other impurities, we guarantee manufacture and quality control practice complying to GMP regulation, as well as all the materials traceable.

Specification

Production according to the following specifications

1. ISO 9001:2015, certified facility.

2. GMP appendix – cell therapy products, State Drug Administration.

3. General introduction to human gene therapy – Chinese Pharmacopoeia 2020, National 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.

Unit Definition

At 37℃, pH 7.5, within 1 h, the amount of enzyme required for totally digestion 1μg λDNA (HindIII digestion, none Dcm methylation) is defined as one unit of enzyme activity.

Storage buffer

10mM Tris-HCl; 300mM NaCl; 1mM DTT; 0.1mM EDTA; 0.5mg/ml HAS; 50% Glycerol; pH 7.4 at 25℃.

Storage temperature

-20±5℃.

Package

Reaction System (50μl)

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Form: Lyophilized Powder

Source: Recombinant plant cell

Package Size: 50ug,100ug,1mg

Purpose of use: cell cultrue

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Form: Lyophilized Powder

Source: Recombinant plant cell

Package Size: 50ug,100ug,1mg

Purpose of use: cell cultrue

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Source: Recombinant plant cell

Package Size: 1g,5g,10g,100g

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Description

Benzo Nuclease is a kind of no-specific endonuclease. The Benzo Nuclease genetically engineered endonuclease from Serratia marcescens.

Benzo Nuclease degrades all kinds of DNA and RNA (double stranded, single stranded, linear and circular and supercoil) but without proteolytic activity. It is effective over a wide range of conditions and possesses an exceptionally high specific activity.

Benzo Nuclease completely digests nucleic acids to 5’monophosphate terminated oligonucleotides 2 to 5 bases in length (below the hybridization limit), which is ideal for removal of nucleic acids from recombinant proteins, enabling compliance with FDA guidelines for nucleic acid contamination. The ability of Benzo Nuclease to rapidly hydrolyze nucleic acids makes the enzyme an excellent choice for viscosity reduction to reduce processing time and increase yields of protein. For example, the enzyme is compatible with BugBuster® and PopCulture® Protein Extraction Reagents and can therefore be added along with these reagents to eliminate viscosity and remove nucleic acids from E. coli extracts. The enzyme consists of two subunits of 30 kDa each. It is functional between pH 6 and 10 and from 0-42°C and requires 1-2mM Mg2+ for activation. The enzyme is also active in the presence of ionic and non-ionic detergents, reducing agents, PMSF (1 mM), EDTA (1 mM) and urea (relative activity depends on specific conditions). Activity is inhibited by>150 mM monovalent cations,>100 mM phosphate,>100 mM ammonium sulfate, or>100 mM guanidine HCl.

• Source: Serratia marcescens
• Molecular weight: 27. 9 KD (SDS)
• Purity: ≥90%, (Ultra≥99%)(SDS-PAGE)
• PI: 6. 85
• Optimum pH: 8. 0
• Optimum temperature: 37°C
• Cofactor: 1～10mM Mg2+
• Form: Liquid enzyme is the clear Solution(Contain Glycerin)
• Solid enzyme is the white lyophilized powder.
• Concentration: 1000 U/µl.
• Activity: ≥1000U/μl
• Specific activity: ≥1. 0x106U/mg protein
• Protease: Not detectable
• Microbial limit: aerobe<10CFU/100 KU Yeast and mold: <10CFU/100 KU
• Endotoxin test: (Gel Clot LAL Assay)<0. 25EU/1000U

Storage Buffer

20 mM Tris-Cl(pH 8. 0), 2 mM MgCl2, 2 mM NaCl, 50%Glycerin

Dilution Buffer

20 mM Tris-Cl(pH 8. 0), 2 mM MgCl2, 2 mM NaCl.

Unit definition

One unit is defined as the amount of enzyme that causes a ΔA260 of 1. 0 in 30 minutes, which corresponds to complete digestion of 37 µg DNA.

Application

• Its high intrinsic activity and broad substrate tolerance make the endonuclease an ideal tool in a variety of biotechnological and pharmaceutical applications:
• removal of nucleic acid from protein samples
• Elimination of nucleic acids from recombinant proteins
• Purification of protein fragments from inclusion bodies;
• Sample preparation in western blotting or two-dimensional gel electrophoresis
• Viscosity reduction in protein extracts.

Shipping and Handling

Liquid enzyme: Ship with Blue Ice, Storage at -20°C
Lyophilized powder: Ship at room temperature, storage blow 4°C

Guarantee

2 years

CAS: 9025-65-4

EC: 3.1.30.2

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This product is a GMP grade recombinant Pyrophosphatase, Inorganic (yeast) (PPase, yeast). It can hydrolyze the inorganic pyrophosphatase produced in nucleic acid amplification experiment, promote the reaction balance to shift to the end of product generation, and increase the amount of product.

This product is a recombinant inorganic pyrophosphatase expressed on a large scale in E. coli.Its molecular weight is about 63kd, which conforms to GMP production and quality management standards. All raw and auxiliary materials are traceable.

Specification

• No virus, mycoplasma and other common pathogens
• Heavy metal: ≤ 10 ppm
• Volume activity (containing cCMP / RNase A): ≥ 600 U / mi
• Internal toxicity control: ≤ 10eu / mg
• Host protein residue: ≤ 50 ppm
• No exonuclease, endonuclease and protease
• Host DNA residue: ≤ 100 pg / mg
• No RNase activity, cleavage activity

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This product is a GMP grade recombinant deoxyribonuclease I (DNase I), which can randomly decompose single or double stranded DNA to produce 5′- p-terminal oligonucleotides. DNase I can cut double stranded DNA at will under Mg2+; Under Mn2+ condition, DNase I can cut DNA double strand at the same site to form flat end or stick end with 1-2 nucleotides protruding.

This product is a large-scale expression of recombinant DNase I in E. coli, with a molecular weight of about 39kD, which conforms to GMP production and quality management specifications. All raw and auxiliary materials are traceable.

Specification

• Heavy metal: ≤ 10ppm
• No virus, mycoplasma and other common pathogens
• Internal toxicity control: ≤ 10 EU / mg
• Volume activity(containing cCMP / RNase A): ≥ 40 Ku / ml
• Host protein residue: ≤ 50 ppm
• No RNase activity, cleavage activity
• Host DNA residue: ≤ 100 pg / mg
• There was no nonspecific nuclease activity

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