| Description | Product description:Hotstart Taq DNA Polymerase High concentration is a thermostable TAQ DNA polymerase modified by an aptamer group. Before high temperature heating, the aptamer group combines with Taq enzyme to inhibit the activity of polymerase, avoid non-specific amplification of primer Product description:Hotstart Taq DNA Polymerase High concentration is a thermostable TAQ DNA polymerase modified by an aptamer group. Before high temperature heating, the aptamer group combines with Taq enzyme to inhibit the activity of polymerase, avoid non-specific amplification of primer extension or the production of primer dimer, enhance the specificity, sensitivity and stability of DNA amplification, and can be widely used in conventional PCR, multiple PCR, nested PCR, etc. After heat shock at 95 ℃ for 10 min, the enzyme can recover its activity. In addition, Hotstart Taq DNA Polymerase High concentration did not detect 3 '→ 5' exonase activity, but it has 5 '→ 3' exonase activity, which can be used for fluorescent quantitative PCR detection. Hotstart Taq DNA Polymerase High concentration has no activity at room temperature, which is convenient for the normal temperature operation of PCR experiment.Product content:1. Hotstart Taq DNA Polymerase High-concentration (10 U/ µ l )2. 5 × Hotstart Taq Buffer ( Mg²⁺ Plus)3. Solution I (10 ×)Activity definition: at 74 ℃ for 30 min, the amount of enzyme required for 10 nmol dNTP to be mixed into acid insoluble sediment is defined as one activity unit.usage method:1. Setting of PCR reaction system:a. Dissolve and mix all solutions required for PCR reaction. It shall be placed on the ice bath or in the ice box. It is recommended that reaction PCR liquid be used separately to avoid repeated freezing and thawing.b. Refer to the following table to set up PCR reaction. It is recommended that the PCR reaction system be configured in an ice bath or on an ice box:※ Dosage of template DNA: To ensure the sensitivity of reaction, 25 µ L The system uses the target sequence copied from 10⁴ as the template. Please refer to the following table to calculate the template amount to be added to the PCR system.one µg Moles of DNA from various sourcesFor example, the concentration of purified human genome DNA is 1 µ g/ µ l. The number of copies of a gene in the human genome is 10, and the number of copies per unit volume is:3.0× 10⁵ mol/µg × 1 µg/µl × 10 copy/mol=3.0× 10⁶ copy/µl1× 10⁴ copy/ (3.0× 10⁶ copy/µl)= 1/300 µlThat is, the concentration of 1/300 ul is 1 ug/ µ The human genome DNA of L contains 10⁴ copies of this gene, diluted 300 times and then added 1 µ L to 25 µ L PCR system. To ensure the specificity of the reaction, the final concentration of DNA should be less than 10 ng/ul, and excessive DNA may have smear bands or even no specific bands.c. Use a pipette to gently blow and mix or slightly Vortex and centrifuge at room temperature for several seconds to make the liquid volume concentrate at the bottom of the tube.d. Place each set PCR reaction tube on the PCR instrument to start PCR reaction.2. Setting of PCR reaction parameters:This product is a aptamer modified Hotstart Taq DNA Polymerase High concentration. In order to optimize the amplification efficiency and quantitative accuracy of PCR, it is recommended that the heat shock time be 95 ℃, 10 minutes3. Product packaging:Product composition1KU5KUStorage temperatureHotstart Taq DNA Polymerase (5 U/µl )200µl1ml-20℃5×Hotstart Taq Buffer（Mg2+ Plus）4ml20ml-20℃Solution I (10×)2ml10ml-20℃... Read More | Collagenase NB 1 is chromatographically highly purified; therefore it contains a very high collagenolytic activity. It is largely free from additional enzymatic activities like clostripain, trypsin-like activity and neutral protease, as well as endotoxins.SpecificationsContains chromatographically Collagenase NB 1 is chromatographically highly purified; therefore it contains a very high collagenolytic activity. It is largely free from additional enzymatic activities like clostripain, trypsin-like activity and neutral protease, as well as endotoxins.SpecificationsContains chromatographically highly purified class I and class II collagenase (1).Largely free from clostripain, trypsin-like protease and neutral protease.Vial contains not less than 2000 PZ U collagenase.Activity (Wünsch): ≥ 3.00 U/mgEndotoxin: ≤ 10.0 EU/mg (Ph. Eur.)(Clostridiopeptidase A)EC 3.4.24.3 • Mr ca. 70 000 - 120 000 (collagenases) • CAS [9001-12-1]ApplicationCollagenase NB 1 is, mostly in combination with Neutral Protease NB, suitable for cell isolation from several tissue types.References and DefinitionsUnit definition: Collagenase: 1 U according to Wünsch (2) catalyzes the hydrolysis of 1 µmole 4-phenylazobenzyloxycarbonyl-L-prolyl-L-leucylglycyl-L-prolyl-D-arginine per minute at 25 °C, pH 7.1.Endotoxin: Ph. Eur. (1 Endotoxin Unit is equal to 1 International Unit of a WHO approved reference standard endotoxin (RSE)).References1. Bond, M.D. & van Wart, H.E. (1984) Biochemistry 23, 3077-30912. Wünsch, E. & Heidrich, H.G. (1963) Hoppe-Seyler's Z. Physiol. Chem. 333, 149-51... Read More | The Leuconostoc GPDH exhibits dual coenzyme specificity, namely NAD and NADP (Olive and Levy, Biochem., 6, 730 730, 1967). When assayed under conditions that are optimal for the particular coenzyme, the ratio of observed catalytic activity is NAD/NADP = 1.8 | Inquire | Inorganic pyrophosphates are inevitably produced in the process of mRNA transcription in vitro. These substances have a great inhibitory effect on transcription. Inorganic pyrophosphatase (PPase) can hydrolyze the inorganic pyrophosphates produced in nucleic acid amplification experiments, promote Inorganic pyrophosphates are inevitably produced in the process of mRNA transcription in vitro. These substances have a great inhibitory effect on transcription. Inorganic pyrophosphatase (PPase) can hydrolyze the inorganic pyrophosphates produced in nucleic acid amplification experiments, promote the shift of reaction equilibrium to the product generation end, and increase the amount of products.The molecular weight of PPase (pyrophosphatase, inorganic, inorganic pyrophosphatase) is about 63kd, which can catalyze the hydrolysis of inorganic pyrophosphate to produce orthophosphate: P2O74_+H2O+PPase→2HPO42_. In the nucleic acid amplification experiment, PPase can hydrolyze the inorganic pyrophosphate generated with the reaction to avoid its inhibition on the reaction system. The removal of pyrophosphate can shift the reaction equilibrium to the product generation end.This product is a GMP level recombinant inorganic pyrophosphatase (yeast source) expressed by large-scale fermentation of E. coli. It is produced with raw and auxiliary materials of medicinal specifications, and the host protein residue and nucleic acid residue are strictly controlled. The product production and quality management procedures in line with GMP specifications ensure that the production process and all raw and auxiliary materials can be traced.Quality requirements project standard appearance Clear liquid Visible foreign matter Compliance with regulations PH value 7.5±8.5 activity 98U/ml-102U/ml purity ≥95% Endonuclease residues Degradation of substrate shall not exceed 10% Exonuclease residues Degradation of substrate shall not exceed 10% RNase residue Degradation of substrate shall not exceed 10% Bacterial endotoxin content ≤10EU/ml Exogenous DNA residue ≤100pg/mg Host protein residue ≤50ppm Mycoplasma detection negative Heavy metal residues ≤10ppm Follow the following specifications1. 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 Committee.4. USP chapter <1043>, adjuvant 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 medical products.Product features1. hydrolyze inorganic pyrophosphate.2. DNA synthesis: significantly enhance DNA replication ability.3. RNA synthesis: increase RNA production in in vitro transcription reaction.4. The optimal reaction temperature is 25℃, and the enzyme can be inactivated at 65℃ for 10min.Product usage1. optimize RNA transcription: improve the RNA yield of in vitro transcription reaction.2. remove PPI contamination from reagents for SNP genotyping by pyrophosphate assay.3. promote the synthesis of protein, RNA and DNA.4. catalyze the reaction of PPI + H2O → 2pi.5. ssr-pcr optimization：Improve efficiency and increase DNA production.Activity definitionCatalytic inorganic pyrophosphate formation 1 per minute under standard reaction conditions µ The amount of enzyme required for mol phosphate was defined as 1 active unit.Preservation system20 mM Tris-HCl; 100 mM NaCl; 1 mM DTT; 0.1 mM EDTA; 50% (v/v) Glycerol; pH 8.0。 Storage temperature-20±5 ℃。Matters needing attention1. the enzyme has activity in various reaction buffers. Generally, the enzyme can be directly added in HDA, lamp and other experiments.2. the dosage of the enzyme needs to be optimized in different experiments, usually adjusted at the concentration of 0.05~1u/ml.3. the optimum reaction temperature of the enzyme was 25 ℃, and it was active at 16~37 ℃, and the enzyme could be inactivated at 65 ℃ for 10min.4. cofactor: mg2+ is necessary for enzyme activity... Read More |