Specific primer design and optimization of annealing temperature for amplification gene peroxidase (POD) in Oryza sativa L.

Abstract

. Peroxidase (POD) is an antioxidant enzymatic that has various functions in the plant life cycle, one of which is as a defense against ROS by catalyzing the conversion of H 2 O 2 to water and O 2 . The ability of the POD enzyme activity to regulate the H 2 O 2 content allows the enzyme to defend plants from stress. One of the methods that can be used to amplify the POD gene is quantitative reverse transcription-PCR (qRT-PCR). This method requires several important components, one of which is the primary (forward and reverse). The primer used in gene amplification must be specific for the target gene so that it can recognize and attach to the desired target gene. This study aims to design suitable primers for POD gene amplification using the qRT-PCR technique. Primers are designed using the PrimerQuest tool. The designed primers were then analyzed for their specificity with geneious prime. Then the primer specificity was checked using the BLAST primer. The results of the primary design with the best criteria for POD gene amplification