HS Hotstart Taq DNA Polymerase
For research use only
P1091 (500 U)
10 × HS Hotstart Taq Buffer (Mg2+ plus)
1 ml × 2
dNTP Mix (10 mM each)
HS Hotstart Taq DNA Polymerase (5 U/μl)
This reagent should be kept at -30~15°C.
HS Hotstart Taq DNA Polymerase is a hot-start Taq DNA polymerase produced by mixing Taq antibody with Taq DNA polymerase in optimum proportion. Based on the thermostable properties of Taq antibody, the activity of HS Hotstart Taq DNA Polymerase remains strictly closed at 55°C, minimizing non-specific amplification during the mixing and heating phases of the reaction system. When the reaction is maintained above 30 sec at 95°C, Taq antibody is completely inactivated and the polymerase activity is fully released, ensuring the extremely high amplification sensitivity and specificity of the PCR system. The activation of HS Hotstart Taq DNA Polymerase is not affected by buffer pH, ion strength or other factors. It is suitable for various hot-start PCR and qPCR reactions based on Taq DNA polymerase. It can be used to amplify low-copy genes from complicated templates (genome, cDNA) and is the preferred hot-start Taq DNA polymerase for molecular diagnostic reagents based on PCR/qPCR. The PCR product has 3 '-d A ends and can be cloned to T vector.
Using activated salmon sperm DNA as template/primer, the activity of ingesting 10 nmol total nucleotides as acid insoluble substances within 30 min at 74°C is defined as 1 unit of activity (U).
Detection of closure of activity: in 1 × Champagne Taq Buffer, the reaction was conducted at 65°C for 30 min, and the released activity was < 5%.
Detection of release of activity: in 1 × Champagne Taq Buffer, after heating at 95°C for 30 sec, and the released activity was > 95%.
Exonuclease residue detection: 50 U of this polymerase and 50 pmol ssDNA and dsDNA substrate were incubated at 37°C for 16 h, and the DNA bands did not change after denaturation PAGE.
Detection of endonuclease residue: 50 U of this polymerase and 0.3 μg of pBR322 DNA were incubated at 37°C for 4 h, and the DNA bands of the plasmids were not changed by agarose gel electrophoresis.
RNase residue detection: 50 U of this polymerase and 1 μg of 293 cell RNA were incubated at 37°C for 30 min, and the RNA bands was unchanged by agarose gel electrophoresis.
E.coli DNA residue detection: the nucleic acid residue in 50 U of this polymerase was detected by E. coli gDNA-specific TaqMan qPCR, and the residue of E. coli genome was less than 10 copies.
Functional detection: In the 20 μl PCR system, cDNA corresponding to the total RNA of 1 pg-200 ng HeLa cells was used as the template to amplify 4 different gene loci. The gradient of PCR product yield could be detected by fluorescence after 35 cycles. At a minimum, the corresponding amplification products were obtained from cDNA corresponding to the total RNA of 1 pg HeLa cells.
10 × HS Hotstart Taq Buffer (Mg2+ plus)a
dNTP Mix (10 mM each)
Primer 1 (10 μM)
Primer 2 (10 μM)
HS Hotstart Taq DNA Polymerase (5 U/μl)c
to 50 μl
a. For most PCR reactions, the optimal final concentration of Mg2+ is 1.5-2 mM. Mg2+ with a final concentration of 2 mM has been included in the system. If necessary, the optimal concentration of Mg2+ can be explored with 25 mM MgCl2 at intervals of 0.2-0.5 mM.
b. The optimal reaction concentration of different templates varies, and the table below shows the recommended amount of 50 μl reaction system templates.
Human genome DNA
E.coli genome DNA
c. The amount of enzyme can be adjusted between 0.125-0.5 μl. In general, increasing the amount of enzyme can increase the amplification yield, but it may decrease the specificity.
* When the GC content of the amplified fragment is > 60%, and normal amplification cannot be achieved after optimizing the conditions, PCR Enhancer (DSBio #P9041) is recommended to optimize PCR reaction.
30 sec (initial denaturation)
7 min (final extension)
* The annealing temperature should be adjusted according to the Tm value of the primer, which is generally set to be 3-5°C lower than the Tm value of the primer.