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

Description

         SP6 RNA polymerase is a DNA dependent RNA polymerase that specifically recognizes the SP6 promoter sequence (5′-ATTTAGGTGACACTATAGAAGNG-3′). SP6 RNA polymerase can catalyze the incorporation of NTP of single stranded or double stranded DNA template downstream of SP6 promoter to synthesize RNA complementary to DNA template downstream of SP6 promoter. Linear plasmids containing SP6 promoters or PCR products containing SP6 promoters can be used as templates for RNA synthesis in vitro.

Accessory reagent

Source

This product is expressed by Escherichia coli, and the expression gene is phage SP6 RNA polymerase gene.

Application

1. Preparation of RNA vaccines and drugs;

2. Synthesis of RNA for in vitro translation;

3. Preparation of radiolabeled RNA probes;

4. Synthesis of antisense RNA for experimental study of expression regulation;

5. Synthesis includes mRNA, siRNA, gRNA and other RNA precursors;

6. Synthesis of Capped mRNA with (Cap analog) as primer.

Storage

Store at -20 ℃ for 24 months.

Storage buffer

50mM Tris-HCl(pH7.9), 100mM NaCl, 1mM EDTA, 20mM β- Mercaptoethanol, 0.1% Triton® X-100, 50% glycerin.

1× RNA polymerase reaction buffer composition

40mM Tris HCl (pH7.9), 10mM DTT, 6mM MgCl2, 2mM spermidine.

Unit Definition

100μL system, one unit enzyme is defined as the amount of enzyme required to add 5 nmol of ATP into polynucleotides within 1 hour at 37 ℃.

Enzyme activity test conditions

50μL reaction system, it contains 1× RNA polymerase reaction buffer, 2mM NTP (0.5mM ATP, GTP, CTP, UTP respectively), 10mM DTT, 1μg DNA template with SP6 promoter.

Quality control

SP6 RNA polymerase does not contain other nucleases, such as T7 or T3 RNA polymerase, DNase and RNA enzyme.

Recommended protocol for RNA synthesis in vitro

1. Template preparation: DNA template containing SP6 promoter and target gene (PCR amplification product, or linearized plasmid). If you use enzyme digestion to linearize the plasmid, it is better to use a restriction endonuclease with a flat end (such as EcoR V), or a sticky terminal enzyme with a protruding 5′ end (such as SapI or BspQI).

2. Precautions before experiment: Make sure that the entire experimental environment is free of RNA enzyme pollution. Use a nozzle, centrifuge tube, and ultrapure water that are free of RNA enzyme. Make sure that the entire reaction system is prepared without RNA enzyme pollution. Make sure that the experimental table, pipette gun, electrophoresis equipment, etc. are free of RNA enzyme pollution. Try to wear masks, hats, and gloves.

3. Reaction system configuration (recommended, system optimization recommended)

4. Incubate at 37 ℃ for 2h.

5. Use DNase I to remove template DNA contamination (optional): every 10μL system add 1U DNase I and incubate at 37 ℃ for 30min. DNase I can be inactivated by heating at 65 ℃ for 10min.

Reference

1. Simon Stammen, Franziska Schuller, Sylvia Dietrich, Martin Gamer, Rebekka Biedendieck, Dieter Jahnl (2010). Application of Escherichia coli phage K1E DNA-dependent RNA polymerase for in vitro RNA synthesis and in vivoprotein production in Bacillus megaterium. Appl Microbiol Biotechnol (2010) 88:529–539.

2. W.Tom Stump, Kathleen B. Hall(1993), SP6 RNA polymerase efficiently synthesizes RNA from short double-stranded DNA templates, Nucleic Acids Research, 5480-5484 Nucleic Acids Research, 1993, Vol. 21, No. 23.

3. Jiyun Yoo 1, Changwon Kang (2000), Bacteriophage SP6 RNA polymerase mutants with altered termination efficiency and elongation processivity, Genetic Analysis: Biomolecular Engineering 16 (2000) 191–197.

4. E D Jorgensen, R K Durbin, S S Risman, W T McAllister (1991), Specific contacts between the bacteriophage T3, T7, and SP6 RNA polymerases and their promoters, Volume 12, Issue 7, November 2019, Pages 908-931.

5. Hirokazu Kotaoi, Yukuo Ishizalci, Nobutsugu Hiraoka and AJrira Obayashi(1987),Nucleotide sequence and expression of the cloned gene of bacteriphage SP6 RNA polymerase, Volume 15 Number 6 1987.

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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

Tth DNA Polymerase is a thermostable enzyme that replicates DNA at 74 °C and exhibits a half-life of 20 minutes at 95 °C isolated from eubacterium Thermus thermophilus strain HB8. Tth catalyzes the polymerization of nucleotides into duplex DNA in the 5´→3´ direction in the presence of magnesium and the polymerization of nucleotides into DNA using an RNA template in the 5´→3´ direction in the presence of manganese. The enzyme has a molecular weight of 94 000 daltons as estimated from the predicted amino acid sequence and exhibits 5´→3´ exonuclease activity. Tth is recommended for use in PCR, RT-PCR, reverse transcription and primer extension reactions at elevated temperature.

Recombinant enzyme with both intrinsic transcriptase and thermostable DNA polymerase activity, a convenient solution for single tube RT-PCR. The enzyme tolerates temperatures up to +50°C –in range of +50 to +70°C for the RT reaction, and up to +95°C for the PCR, overcoming problems caused by RNA secondary structures. Carryover prevention via the incorporation of dUTP and subsequent treatment with UNG is also possible.

Features

The thermostability and the reverse transcriptase (RT) activity of Tth DNA polymerase is useful in amplifying DNA from RNA templates that contain G-C-rich sequences or secondary structures since the elevated temperatures serve to denature the template RNA. Higher temperatures (in contrast to other enzymes for RT-PCR) also result in increased specificity of primer hybridization and extension. The concentration of RNA template for effective reverse transcription with Tth DNA polymerase should be higher if to compare with reverse transcription directed by Reverse Transcriptase (M-MuLV, AMV).

Applications

• PCR and RT-PCR – cDNA synthesis

Concentration: 5 u/µl

Storage Buffer

10 mM Tris-HCl, 1 mM dithiothreitol, 0.1 mM EDTA, 300 mM KCl, 0.1% Triton X-100 (v/v)*, 50% glycerol (v/v), pH 7.5 (25°C)

Reaction Buffer

5X RT/PCR reaction buffer (One Step-buffer): 250 mM bicine (pH 8.2, by KOH, at 25°C), 580mM KOAC, 40% Glycerol
10X PCR buffer: 100 mM Tris-HCl, (pH 8.8 at 25°C), 15 mM MgSO₄, 800mM (NH4)₂SO₄, 0.5 mg/ml BSA, 0.5% Tween 20

Usage

1.) One step RT PCR: – Reverse transcription and amplification in one Tube – Advantage: The one step One step reaction eliminates the risk of cross contaminations associated with two step RT-PCR.
2.) Two step RT PCR
3.) Standard PCR

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Description

Taq DNA polymerase is isolated from the E. coli cloning Thermus aquaticus. The molecular weight of the product is approximately 94 kDa. EasyTaq DNA polymerase has the 5′ to 3′ DNA polymerase activity and 5′ to 3′ exonuclease activity without 3′ -5′ exonuclease activity. The extending speed is 1-2 kb/min. There is an “A” on 3′ end. The PCR product can be cloned in TA vector.

• CAS No. : 9012-90-2
• Other Names: nonstandard Taq DNA Polymerase
• Purity: 99.9%
• Classification: Specific Reagents
• nonstandard Taq DNA Polymerase: High concentration 100Unit

Package Details

• small package or bulk package both OK. Ship with blue ice and form box.
• storage buffer without glycerin.
• Offer OEM ODM and bulk service!
• high-sensitivity, high amplification Taq DNA polymerase. offer to all over the world, supply Bulk and OEM.

Characteristics

(1) high-sensitivity
(2) high amplification efficiency

Unit Definition

One unit of Platinum Taq DNA Polymerase High Fidelity incorporates 10 nmol of deoxyribonucleotide into acid-precipitable material in 30 minutes at 74°C.

Quality Control

• functional absence of double- and single-stranded endonuclease activity;
• Purity>90% homogeneous by SDS gel electrophoresis.
• Each lot of EasyTaq DNA Polymerase is assayed for amplification from as little as 10 ng of human genomic DNA.

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Description

The Hot Start Taq DNA Polymerase is a recombinant Taq DNA polymerase which has been chemically modified by the addition of heat-labile blocking groups to its amino acid residues. The enzyme is inactive at room temperature, avoiding extension of non-specifically annealed primers or primer dimers and providing higher specificity of DNA amplification. The functional activity of the enzyme need to be activated in incubation at 95°C for 10 minute. The activated enzyme maintains the same functionality.

Taq DNA polymerase: Has the 5′ to 3′ DNA polymerase activity and 5′ to 3′ exonuclease activity without 3′ – 5′ exonuclease activity. The extending speed is 1 kb/30 sec. There is an “A” on 3′ end. The PCR product can be cloned in TA vector.

Applications

• Hot-start PCR amplification
• Specific amplification of complex cDNA and genomic template
• Amplification from low copy number DNA template
• Real-Time PCR
• Multiple PCR
• Generation of PCR products for TA cloning, Gene microarray analysis and SNP test

Quality Control

• Functional absence of double and single-stranded endonuclease activity;
• Purity>99% test by SDS gel electrophoresis;
• Each lot of HotStar Taq DNA Polymerase is assayed for amplification from as little as 10 ng of human genomic DNA;
• Retain full activity at room temperature for one week;
• No host DNA residue.

Concentration: 5 u/μl

Package: Bulk

Storage: Store at -20°C

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Product Number: PC09

Shipping and Storage

-20°C.

Components

Note: The 5×Golden Star Taq PCR Buffer of this product contains 8.5mM magnesium ions.

Description

Golden Star Taq DNA Polymerase is a chemically modified new and efficient Taq DNA Polymerase, which is completely blocked at room temperature, making the enzyme inactiveat low or normal temperature. Thus, non-specific amplification caused by non-specific binding of primers and templates or primer dimers can be effectively avoided at room temperature ,the activation of the enzyme must be incubated at 95°C for 10 minutes.The unique buffer system enables the enzyme to be widely used, and makes efficient amplification of templates with high GC content, complex secondary structure and low copy. Using this product for PCR amplification, the 3′ end of the PCR product with an “A” base, can be directly used for T/A cloning.This product has strong specificity and can be directly used for downstream cloning or chip hybridization experiments without the need for glue recovery to remove the heteroband after PCR amplification. It is mainly for conventional PCR, RT-PCR, real-time PCR, multiplex PCR, gene chip analysis and SNP detection, especially for PCR reaction with high specificity requirements.

Activity Definition

Using activated salmon sperm DNA as template/primer, the amount of enzyme required to incorporate 10nmol deoxynucleotides into acidic insoluble material was defined as 1 active unit(U) at 74°C for 30 min.

Quality Control

The purity of SDS-PAGE was greater than 99%. No exogenous nuclease activity was detected. No host residual DNA was detected by PCR. It can effectively amplify single copy genes in the human genome. There was no obvious activity change after one month storage at room temperature

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LA Taq DNA Polymerase three times higher than Taq DNA Polymerase

Long PCR Polymerase which synergistically generate long PCR products with greater yield and fidelity than Taq DNA Polymerase alone.

The fidelity of PCR is three times higher than with Taq DNA Polymerase.

The LA Taq is optimized for generation of very long amplicons: up to 40 kb with viral DNA and up to 15 kb with genomic DNA templates. The specially formulated LA Taq Buffer protects DNA from depurination and nicking during long thermal cycling.

The PCR products generated with the LA Taq DNA Polymerase are mostly 3′-dA tailed, which can be cloned in TA vector.

Features

• Long PCR products
• up to 40 kb with viral DNA as template
• up to 15 kb with genomic DNA as template
• Ideal for GC-rich templates up to 85% GC.
• Fidelity is three-times higher than with Taq DNA Polymerase.
• High yields. Incorporates modified nucleotides.

Quality Control

Functionally tested in generation of 40 kb amplicon with lambda DNA as template.

Concentration: 5 u/μl

Storage

Storage Buffer: The enzyme is supplied in: 20mM Tris-HCl (pH 8.0); 0.1mM EDTA; 1mM DTT; 100mM KCl; Stabilizers; 50% glycerol.

Storage: Store at -20°C.

Reaction

Reaction PCR Mixture Set Up and Recommended thermal cycling conditions: The flowing is only an example which take a 20KB human genomic DNA as the template, that only for a reference, It can be adjusted according the length and the sequence of the template and the primer.

94°C 3 min
94°C 30 sec
55°C 30 sec 30 cycles
72°C 1 min
72°C 5 min.

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Description

Taq Plus DNA Polymerase is a kind of High Fidelity Taq DNA Polymerase. mixture of Taq DNA Polymerase and proofreading DNA Polymerase, which allows for the amplification of long templates, up to 30kb, with high fidelity. The two enzymes act synergistically during PCR to generate more accurate and longer PCR products with greater yields compared to Taq DNA Polymerase alone. PCR products, amplified up to 10kb in length with Taq Plus DNA Polymerase, generate a mixture of blunt ends and single base (A) 3′ overhang. The products can be used for direct T/A cloning, but its efficiency is not as high as PCR products amplified with Taq polymerase alone.

Taq Plus DNA Polymerase is a special formulation designed for amplify large fragment. The main component is Taq DNA Polymerase, and Pfu DNA Polymerase are added to enhance the efficiency of amplification reaction. Theoretically, Taq Plus produces significantly higher yields of PCR products than ordinary Taq Polymerase, especially for fragments >1kb, and can amplify up to 20kb. Taq Plus also contains a proofreading activity that reduces the error rate of Taq Polymerase. Most of the amplified DNA fragments have a 3´A overhanging. However, a small percentage of the amplified DNA fragments are blunt-ended. Taq Plus is suitable as a direct replacement for ordinary Taq Polymerase in most applications.

Application

Long PCR (up to 20 kb), PCR cloning, RT-PCR etc.

Features

High fidelity: with an error frequency of 1.6X10-6 during DNA synthesis.

Higher yield: Taq Plus increases the efficiency of polymerization reaction, resulting in a great percentage of extenuation reaction completion up to 20kb

Unit Definition

One unit of the enzyme catalyzes the incorporation of 10nmole of deoxyribonucleotides into a polynucleotide fraction in 30min at 74°C

Quality Control

No contaminating endonuclease or exonuclease activity detected. Functionally tested in PCR.

Concentration: 5u/µl

Storage

Storage Buffer: 20mM Tris-HCl, pH 8.0, 100mM KCl, 0.1mM EDTA, 5mM DTT, 50% glycerol, 0.5% NP40 and 0.5% Tween 20.

Storage: Store at -20°C

10X Reaction Buffer (Mg²⁺ Plus)

500mM KCl, 100mM Tris-HCl (pH 9.0 at 25°C) and 1% Triton X-100, 100mM (NH4)₂SO₄, 15mM MgCl2, PCR enhancer

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