Optimization of PCR method for the polymorphic detection of beta-2 adrenergic receptor gene / Afriza Aini Mohamad Afzal

• Main Author: Mohamad Afzal, Afriza Aini
• Format: Thesis
• Language: English
• Published: 2006
• Subjects: RM Therapeutics. Pharmacology; RS Pharmacy and materia medica
• Online Access: https://ir.uitm.edu.my/id/eprint/99089/1/99089.PDF; https://ir.uitm.edu.my/id/eprint/99089/

The aim of this study is to perform a one step direct PCR method in identifying polymorphisms of five alleles of interest of the beta-2 adrenoceptor gene (Arg 16/ Gly 16, -20T/C, Gln27/ Glu27, -47T/C and Thr164/ Ile164). This one step direct PCR is also expected to produce better results compared to a previous study with the same alleles of interest but using a different method which was the two steps PCR. The method used in the study was multiple allele specific reaction which can allow simultaneous amplification of the five beta-2 adrenoceptor alleles. Specific primers were used to complement the target sequence and produce specific bands of interest. The PCR method was optimized for reproducibility and specificity. Few parameters were frequently adjusted to produce the desired results. The parameters include annealing temperature, cycling conditions, primers and Taq polymerase concentration. At the end of the study, the results were unsuccessful due to few problems and possible causes that can be improved in future studies.