Hieff® Taq DNA polymerase is a thermostable recombinant DNA polymerase expressed by Thermus aquaticus with a molecular weight of 94 KD. It has 5'→3' polymerase activity and 5'→3' exonuclease activity, but no 3'→5' exonuclease activity. The amplified product has 3'- dA and can be directly used for TA cloning.
10×Taq Buffer (Mg2+ Free)
25 mM MgCl2
Genotyping, colony PCR and other conventional PCR.
Using the activated DNA of salmon sperm as template / primer, the activity was defined as one active unit (U) when 10 nmol of total nucleotide was ingested as acid insoluble substance at 74℃ for 30 min.
Exonuclease Activity: In 20 μL reaction system, 10 U of this product and 0.6 μg of λ - HindIII were incubated at 74℃ for 1 h, and the electrophoresis band of DNA had no change.
Endonuclease Activity: In 20 μL reaction system, 10 U of the product and 0.6 μg of supercoiled pBR322 DNA were incubated at 74 ℃ for 1 h, and the DNA electrophoretic bands did not change.
Residual E.coli gDNA Test: The E.coil 16S rDNA gene was amplified by adding 2 U of the product into 50 μL system and using sterile ddH2O as template. After 35 cycles, the amplified products were subjected to 1% agarose gel electrophoresis and EB staining without amplification bands.
Transport in ice packs，it can be stored at -20℃ for 2 years.
For your safety and health, please wear experimental clothes and disposable gloves.
10×Taq Buffer(Mg2+ Free)
25 mM MgCl2
dNTP Mix (10 mM each)
Forward primer (10 μM)
Reverse primer (10 μM)
Hieff® Taq DNA Polymerase (5 U/μL)
Final concentration of Mg2+：The best concentration of Mg2+ is 1.5-2 mm. If necessary, the best concentration of Mg2+ can be found at intervals of 0.2-0.5 mm.
Polymerase addition: at room temperature, the polymerase has a certain degree of 5'- 3' polymerase activity. In order to prevent non-specific amplification, it is suggested to add the polymerase to the reaction system in the last step.
Concentration of polymerase：0.04 U/μL is recommended. It can be optimized between 0.025-0.04 U/ μL.
Recommended use of different templates (50 μL reaction system)：
Type of template
50 ng-100 ng
10 pg-20 ng
1-5 μL (No more than 1/10 of the reaction system)
30 sec-5 min
Pre denaturation temperature and time：94℃ is recommended. The recommended pre denaturation time is 30 sec for plasmid DNA and other simple templates; 3 min for complex templates such as cDNA and genomic DNA; The template with high GC was 5-10 min.
Annealing temperature and time: 60℃ is recommended. Temperature gradient can be set up to find the optimum temperature of primer annealing. The recommended annealing time is set to 20 sec and can be adjusted in 10-30 sec. The annealing time is too long, which may lead to the diffusion of the product on the adhesive.
Amplification products: Please store the PCR amplification products at - 20℃ to prevent DNA degradation.