Hot Start Taq DNA Polymerase (C09-0912-087)

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℃. Specifications and Purity: 5U/μL. Related Document: https://ald-pub-files.oss-cn-shanghai.aliyuncs.com/aladdinsci/pdp/sds/1/H292167-SCI_73b6f21d55f8c17d19078d66c45f6dfc.pdf.