Heat shock condition: 95 ℃, 2.5min
Externase activity: 5 '→ 3'
Matching of pollution prevention system: √
ARMS: √
SNP: √
Fluorescence quantitative PCR: √
Additional AT carrier connection added: √
Recommended items: qualitative PCR, quantitative PCR Product manual Anstart Taq II DNA polymerase is a mixed product of anti-Taq monoclonal antibody and Taq II DNA polymerase. It is an upgraded version of the original Anstart Taq DNA Polymerase (MD006) product, with better stability and specificity. Before heating at high temperature, the anti-Taq monoclonal antibody binds to Taq polymerase to inhibit the activity of the polymerase, thereby inhibiting non-specific annealing of primers or non-specific amplification caused by primer dimers under low temperature conditions. When the amplification reaction system is heated to 95°C for 2 minutes, the polymerase activity is restored due to the denaturation of the anti-Taq monoclonal antibody, so there is no need for special inactivation treatment, and it can be used under conventional PCR reaction conditions. By using it in conjunction with an improved buffer, rapid activation can effectively increase the amount of reaction products and improve the sensitivity and specificity of the PCR reaction. It can be applied to hot-start PCR to amplify complex templates and low-copy target fragments. Product content
1. Anstart Taq II DNA Polymerase ( 5 U/μl ) 2. 10×Anstart Taq II Buffer (without Mg2+ ) 3. 100mM MgCl2 Product Usage The hot start method is used for PCR amplification. Instructions 1. PCR reaction system settings a. Dissolve and mix the various solutions required for the PCR reaction, and place them on an ice bath or in an ice box. It is recommended to use aliquots of the reaction PCR liquid to avoid repeated freezing and thawing. b. Refer to the following table to set up the PCR reaction. It is recommended to configure the PCR reaction system in an ice bath or on an ice box: Reagent | Volume | Final concentration | 5×Anstart Taq II Buffer (Mg2+ Plus) | 10μl | 1× | dNTP（25mM each） | 0.4μl | 0.2mM | Template DNA | 10μl | — | Primer I（10μM） | 1μl | 0.2μM | Primer II（10μM） | 1μl | 0.2μM | Anstart Taq II DNA Polymerase ( 5 U/μl ) | 0.5μl | — | Ultra-pure water | Up to 50μl | — | Total capacity | 50μl | — |
The recommended dosage for different types of templates in a 50μl reaction volume is as follows: Mammalian genomic DNA: 0.1-1μg Escherichia coli genomic DNA: 10-100ng Plasmid DNA: 0.1-10ng Too much template DNA can easily lead to non-specific PCR products c. Use a pipette to mix gently or gently Vortex to mix, and centrifuge for a few seconds at room temperature to allow the liquid to accumulate at the bottom of the tube. d. Place each set of PCR reaction tubes on the PCR machine to start the PCR reaction. 2. The setting of PCR reaction parameters takes the amplification of 1Kb target fragment as an example Step | Temperature | Time | Number of cycles | Predenaturation | 95℃ | 2.5min | 1 | Transsexual | 94℃ | 30s | 25-35 | Annealing | 55℃ | 30s | Extend | 72℃ | 1min | Last extension | 72℃ | 10min | 1 |
a. PCR reaction settings need to be based on the template, primers, PCR product length and GC content and other conditions to set different PCR reaction conditions, including temperature, time and number of cycles. b. The time setting of STEP4 (extension) needs to be set according to the length of the PCR product, usually the extension time per kb product is 1 min. For example, if the length of the PCR product is 1kb, the extension time can be set to 1min, and the length of the PCR product is 2kb, then the extension time can be set to 2min, and so on. c. For the first PCR, in order to ensure that the expected PCR product can be amplified as much as possible, the number of cycles can be set to 35. The number of PCR reaction cycles that need to be semi-quantitative or quantitative must be appropriately optimized to make the PCR reaction reach the plateau. | 
