Biochemical Techniques and Instrumentation

6.1.3. Components of PCR

A basic PCR set up requires the following components and reagents:

• DNA template: This contains the specific DNA target region to be amplified.
• Primers: Two primers that are complementary to the 3’ ends of each of the sense and anti-sense strand of the DNA target.
• Taq polymerase: This is a DNA polymerase with a temperature optimum at around 70 °C.
• Deoxynucleoside triphoshates (dNTPs): They are the building blocks from which DNA polymerases synthesize a new DNA strand.
• Buffer solution: This provides a suitable chemical environment for optimum activity and stability of the DNA polymerase.
• Divalent cations: Magnesium ions (Mg²⁺) is generally used. Sometimes manganese ions (Mn²⁺) are utilized for PCR-mediated DNA mutagenesis.
• Monovalent cation: Potassium ions