GENERAL KNOWLEDGE

KEY FEATURES REQUIRED FOR A DNA POLYMERASE FOR POLYMERASE CHAIN REACTION

DNA polymerase is a key enzyme used in the polymerase chain reaction (PCR) process, which is essential for amplifying specific DNA sequences. The key features required for a DNA polymerase to be suitable for PCR include:

  1. Thermal Stability: The DNA polymerase should be able to withstand the high temperatures used in PCR, typically around 95°C during the denaturation step. This thermal stability ensures that the enzyme remains active throughout the PCR cycles.
  2. Processivity: A high processivity is crucial for efficient DNA synthesis during PCR. Processivity refers to the ability of the polymerase to add multiple nucleotides to a growing DNA strand without dissociating from the template.
  3. Fidelity: The DNA polymerase should have a high fidelity, meaning it accurately incorporates nucleotides and has proofreading capabilities to minimize errors during DNA synthesis. This is particularly important for generating accurate PCR products.
  4. Amplification Efficiency: The polymerase should efficiently amplify the target DNA sequence without introducing biases or artifacts, ensuring reliable and reproducible results.
  5. Lack of Exonuclease Activity: The DNA polymerase should not possess 3′-5′ exonuclease activity, as this could lead to degradation of primers and non-specific cleavage of DNA molecules.
  6. Compatibility with PCR Buffer Conditions: The polymerase should be compatible with the buffer conditions used in PCR, including optimal pH and salt concentrations.
  7. Inhibitor Resistance: The enzyme should be resistant to common PCR inhibitors present in complex biological samples, allowing for successful amplification from various sample types.
  8. Commercial Availability: Availability from commercial sources with consistent quality and performance is also an important practical consideration.

Several DNA polymerases have been developed or engineered to meet these requirements, with Taq polymerase being one of the most well-known and widely used in PCR due to its robustness and processivity at high temperatures. Other examples include Pfu (Pyrococcus furiosus) polymerase known for its high fidelity and proofreading activity, and various engineered polymerases with enhanced properties tailored for specific PCR applications.

In summary, a DNA polymerase suitable for PCR should exhibit thermal stability, high processivity, fidelity, amplification efficiency, lack of exonuclease activity, compatibility with PCR buffer conditions, inhibitor resistance, and commercial availability.

Hearing god’s voice in unlikely places : aaron watson & anthony lucia.