Question

What are the important components for a successful Polymerase Chain Reaction (PCR)?

Answer 1

Polymerase Chain Reaction (PCR) requires following important components:

1.Thermostable DNA Polymerase:

DNA polymerases are required for amplification of target DNA. Taq polymerase, a DNA polymerase functional at high temperatures above 75° C is used in routine PCRs. 

2. Primer pair: A pair of short synthetic oligonucleotides are required to prime DNA synthesis. There specificity is crucial for high efficiency. 

3. Deoxyribonucleoside triphosphates (dNTPs): Equimolar amounts of adenine (A), guanine (G), cytosine (C), and thymine (T) dNTPs serve as  building blocks for amplified DNA copies.

4. Divalent Cations: Normally Mg2+ ions (MgCl₂) are needed for enzyme activity, a  cofactor function. 

5. Buffer: A reaction buffer (Tris-cl) is used for pH maintenance which is crucial for enzymatic reactions taking place. 

6. Monovalent Cation: K+ from KCl helps facilitate primer annealing. 

7.Template DNA: Specific target DNA sequence serves as template DNA for the ampilification process. Single or double stranded DNA type can be used. 

 

Polymerase Chain Reaction (PCR) requires following important components: DNA template, DNA polymerase enzyme , primers, nucleotides, cofactor ions and reaction buffers.