DNA Extraction and Characterization of Salmonella's Lytic Bacteriophage Genome Pembimbing

Abstract

Bacteriophages are infectious virus that can replicate only inside their specific bacterial host. Bacteriophage can be used as a biocontrol agent for a specific bacteria such as Salmonella and other applications. Understanding of their molecular genetics is important due to their potential applications. The objectives of this research was to study three isolates of Salmonella‟s phages and to find out the best method to isolate and characterize genomic DNA of the phages. Three different methods i.e. PEG/NaCl Method, Scraping plaque method and Enrichment Method were used to concentrate phages for genomic DNA extraction. Quantity and purity of extracted DNA was determine based on their optical density ratio of 260/280 nm and agarose gel electrophoresis. Genomic DNA size of the phages were determined using agarose gel electrophoresis with High range ladder as a marker. The best method to prepare bacteriophage concentrates for DNA genome extraction was enrichment method. This method had higher concentration and purity of extracted DNA. Based on DNA genome size, the bacteriophage genome of Salmonella sp were comparable with other study of Salmonella phages. Phage P15 with genome size of 39,5 kb was close to phage epsilon15. Phage P19 with genome size of 41 kb was close to phage ST160, and phage P38 with genome size of 48,5 kb was close to phage Gifsy-1.