Nurhayati, Siti2021-07-292021-07-292020-08wahyu sari yenihttps://repository.unri.ac.id/handle/123456789/10079Pandan (Benstonea sp.) is one of the most important plant in Kajuik Lake located in Langgam, Riau Province, Indonesia. Polymerase Chain Reaction (PCR) is a technique used to multiply or amplify DNA samples in vitro using a primer pair. trnQ-5’ rps16 intergenic spacer is a pair of primers used to amplify the trnQ- 5’ rps16 intergenic spacer region. This study aimed to optimize annealing temperature of trnQ-5’ rps16 intergenic spacer primer. This research invloved total DNA isolation using Genomic DNA Mini Kit Plant (Geneaid), PCR, and electrophoresis. The result showed that the optimal annealing temperature for amplification of trnQ-5’ rps16 intergenic spacer was 58°C. The PCR product on this annealing temperature was thick DNA with a size of about 950 bp .enAnnealingBenstonea spPCRArticle