| 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 | Inquire | Malic Dehydrogenase is a ubiquitous enzyme, which exists in two isoforms in eukaryotic cells.Malic dehydrogenase exists as a dimer with each subunit containing an NAD-binding domain and a substrate-binding carboxy-terminal domain required for activity. Malic dehydrogenase is a cytoplasmic isozyme Malic Dehydrogenase is a ubiquitous enzyme, which exists in two isoforms in eukaryotic cells.Malic dehydrogenase exists as a dimer with each subunit containing an NAD-binding domain and a substrate-binding carboxy-terminal domain required for activity. Malic dehydrogenase is a cytoplasmic isozyme and an important catalyst in the tricarboxylic acid cycle.ReagentsA. 0.1 M Tris-HCl buffer (pH7.8)B. 0.01 M Phosphate buffer (KH2PO4-NaOH, pH 7.0)C. Triton X-100 solution (50 mg/ml)D. 0.01 M Phosphate buffer containing 0.1% Triton X-100 (KH2PO4-NaOH, pH 7.0)Dilute 20 ml of Triton X-100 solution (C) with approx. 800 ml of 0.01M Phosphate buffer (B). Fill up to 1,000 ml with 0.01M Phosphate buffer (B).E. NADH soluton Weigh 9 mg of NADH and dissolve in 0.1M Tris-HCl bufer (A). Fill up to 50 ml with 0.1M Tris-HCl Buffer (A). (Can be used for 5 days if kept refrigerated)F. Substrate solutionWeigh 11 mg of oxaloacetic acid and dissolve in 0.1M Tris-HCl buffer (A). Fill up to 50 ml with 0.1M Tris-HCl buffer (A) (Make a fresh solution for each use.)G. Enzyme solutionWeigh out Malate Dehydrogenase and dissolve in chilled 0.01M Phosphate Bufer containing 0.1% Triton X-100 (D). Enzyme solution should be prepared so that the value of AOD/minute becomes in the range of 0.025 ± 0.010.ProcedurePipette 2.0 ml of NADH solution (E) and 0.90 ml of Substrate solution (F) respectively into a quartz cell (d=10 mm) and keep at 25 + 0.5'℃ for 5 minutes. Then, pipete 0.10 ml of Enzyme solution (G) into the quartz cell and mix well immediately. Keep the reaction mixture at 25 ±0.5'C.Exaclly at 2 minutes and 5 minutes after the addition of Enzyme solution (G), measure the absorbances of the reaction mixture at 340 nm(A2 and A5).As a blank, pipette 0.01M Phosphate buffer (D) into another quartz cel (d=10 mm) instead of the Enzyme solution (G) and follow the same procedure described above (Ab2 and Ab5).CalculationMalate dehydrogenase activity (u/mg)=[(A2-A5)-(Ab2-Ab5)]/3*(1/6.22)*(n/0.1) ApplicationThis enzyme is used for the enzymatic determination of L-malate and gluamate oxalo-acetate transaminase(GOT)in clinical diagnosis... Read More | 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 | Purity>95% (SDS-PAGE&HPLC) Endotoxin level<0.1 EU/µgFunctionMay regulate apoptosis, cell proliferation and cell differentiation. Binds beta-galactoside and a wide array of complex carbohydrates. Inhibits CD45 protein phosphatase activity and therefore the dephosphorylation of Lyn Purity>95% (SDS-PAGE&HPLC) Endotoxin level<0.1 EU/µgFunctionMay regulate apoptosis, cell proliferation and cell differentiation. Binds beta-galactoside and a wide array of complex carbohydrates. Inhibits CD45 protein phosphatase activity and therefore the dephosphorylation of Lyn kinase.Gal-1 is also engaged in many protein-protein interactions. Gal-1 plays a number of crucial roles in neuronal cell differentiation and survival in both the central and the peripheral nervous systems, and the establishment and maintenance of T-cell tolerance and homeostasis in vivo... Read More |