Study of Factors Affecting Gene Amplification in Polymerase Chain Reaction

Background and Technology has caused great progress in the novel molecular diagnosis and research methods in medical laboratory. Novel methods have also led to the higher accuracy rates in laboratory procedures which are of considerable importance in clinical follow-up of genetic diseases. Therefore this study is intended to investigate factors affecting gene amplification in polymerase chain reaction in order to enhance diagnostic accuracy. This deh1ive analytical research was conducted on 61 adenocarcinoma specimens in the cellular and molecular departments of Sabzevar and Isfahan Universities of Medical Sciences in Iran. DNA was extracted by the standard kit; then the segment AURKA gene and P53 Gene were amplified using two pairs of specific primers and different concentration Mgcl2 in a PCR assay. PCR product was electrophoresized in agarose gel. Electrophoresis of PCR product with Mgcl2 concentrations of 3 and 5 mm was better than 1.5 mm. The primer with concentration of 1 mm was better than 5 and 10 mm. From the two pairs of primers used in amplifying AURKA gene axon 4 one pair of primers was better than the other pair. From the two pairs of primers used for amplifying the axon 5 of P53 gene in PCR assay one pair was better than the other. Primer type and concentration of Mgcl2 are important in amplifying genes in the polymerase chain reaction assay.