Basic Principles Of The Polymerase Chain Reaction ( PCR )

Decent Essays
a. Describe the basic principles of the polymerase chain reaction (PCR).
PCR is biochemical research technique developed in 1985 by Kary Mullins. It is based on the principle of amplifying DNA through a series of cycling reactions with varying temperatures. PCR uses the ability of thermostable polymerase enzymes to synthesize a new complementary DNA strand to a template strand.

b. What are the essential components of a PCR reaction?
The essential components of PCR reactions are:
i. Taq polymerase: A thermostable DNA polymerase isolated from the bacterium Thermus aquaticus that survives at 750C. ii. DNA primers: Approximately 20 nucleotides in length that are specific to a DNA region of interest. They are complementary in sequence to the ends of the target DNA. Primer is chosen based on what part of DNA needs to be copied or amplified. iii. Sample DNA: Containing target DNA iv. Nucleotides: dNTPs or deoxynucleotide triphosphates are single units of bases of A, T, G and C.
v. PCR Buffer: Contains Tris-HCL, KCL, and MgCl2. Buffer helps to create optimal conditions for the activity of Taq DNA polymerase.

c. What is the name of the instrument typically used for PCR? Describe its operations and how it is useful for performing PCR reactions.
PCR reaction is typically performed in a thermocycler also called as a PCR machine or DNA amplifier.

Thermocycler operation and PCR process:
Thermocycler consist of a thermal block with holes to fit tubes in which the PCR reaction
Get Access