Potential Panacea to the Complexities of Polymerase Chain Reaction (PCR)

Abstract

The polymerase chain reaction (PCR) allows the exponential amplification of target DNA sequences, and has greatly impacted the world of scientific research. Although the reagents, equipment needed and the process (denaturation, annealing and elongation) appears simple and straightforward, PCR is adorned by complexities. Hence this paper seeks to discuss the complexities often encountered whilst performing PCR. Findings showed that primers, annealing temperature, wholesomeness and purity of template DNA, PCR contaminants and inhibitory substances, choice of polymerase, Magnessium chloride concentration, and number of cycles influence the outcome of a PCR. These factors sometimes enhance quality but compromise yield and vice versa. A high amount of Magnesium chloride or number of cycles in a PCR would result to a good yield but compromises quality and vice versa. In contrast, a high quality but lower yield of PCR is attained where the annealing temperature is either close to or higher than the melting temperatures of the primers.  Thus performing PCR is more or less choosing between quality and yield, and this paper would enable researchers strike the needed balance to overcome some of the associated complexities.

Keywords: PCR, DNA, Taq polymerase, PCR inhibitory substances, primers