Optimisation of Standard PCR Programme

Abstract

PCR is a very sensitive technique that gained its significance through the high contribution ona variety of bioscience fields. It is not surprising that the most cited scientific paper inbiosciences is related to the PCR programme. The optimisation of the PCR is very importantto achieve amplification with reliable results. Variables like Mg++ concentration, templateDNA dilution, annealing temperature, and primers' concentration were optimized usingclassical and touchdown amplification methods. Other PCR methods like hot-start canimprove the quality and the quantity of the PCR yield. The utilization of mathematical andstatistical ways in designing a PCR programme like Taguchi or chessboarding methods canefficiently enhance the PCR reaction. Also the usage of very pure reagents and the utilizationof a suitable kind of the Taq polymerase would highly improve future experiments. This workwas performed to find the optimal reaction conditions for the utilized standard PCRprogramme.