**Background**
The polymerase chain reaction (PCR) has revolutionized molecular biology by allowing the exponential amplification of specific DNA sequences. Central to this process is the requirement for a DNA polymerase that can withstand the high temperatures necessary for DNA denaturation. Thermostable polymerases, derived from thermophilic organisms, are essential for this cycle, enabling the precise replication of genetic material for applications ranging from genetic diagnostics to forensic analysis. In the context of modern biotechnology, enhancing the robustness of these enzymes to handle complex templates, such as those found in clinical samples, is of significant research importance. Therefore, we will introduce a modified thermostable enzyme – Taq DNA polymerase.
**Definition**
Taq DNA polymerase is a thermostable DNA polymerase with an enzyme activity of ≥50 U/μL, where one unit is defined as the amount of enzyme that catalyzes the incorporation of 1 nmol AMP into the polynucleotide at 37°C in 60 min.
**In Vitro Studies**
According to the Taq DNA polymerase description, this product is produced by modifying the protein molecule of wild-type Taq DNA polymerase and expressing it in E. coli to enhance the enzyme’s anti-interference ability. The Taq DNA polymerase in vitro performance is particularly effective for PCR amplification directly from whole blood. Following the Taq DNA polymerase protocol, a typical 25 μL reaction system consists of 2.5 μL reaction buffer (100 mM Tris, 500 mM KCl, 15 mM MgCl2, 0.8% NP-40, pH 8.8), up to 5 μL whole blood DNA template, 2.5 μL 2.5 mM dNTP, 1 μL each of upstream and downstream primers (10 μM), and 0.5 μL Taq DNA polymerase. The thermal cycling conditions include an initial denaturation at 95°C for 3 min, followed by 35-40 cycles of 94°C for 30 sec, 55°C for 30 sec, and 72°C for 45-90 sec, with a final extension at 72°C for 7 min. In conclusion, Taq DNA polymerase is a highly stable and modified enzyme suitable for the amplification of DNA from challenging biological samples.
Keywords
Taq DNA polymerase, 2304873-37-6, DNA/RNA Synthesis, Inhibitor, inhibitor, inhibit
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