title: Efficiency comparison of three methods for extracting genomic DNA of the pathogenic oomycete Pythium insidiosum.
abstract: BACKGROUND
  The fungus-like organism Pythium insidiosum is the causative agent of a life-threatening tropical infectious disease, pythiosis, which has high rates of morbidity and mortality. A lack of reliable diagnostic tools and effective treatments for pythiosis presents a major challenge to healthcare professionals. Unfortunately, surgical removal of infected organs remains the default treatment for pythiosis. P. insidiosum is an understudied organism. In-depth study of the pathogen at the molecular level could lead to better means of infection control High quality genomic DNA (gDNA) is needed for molecular biology-based research and application development, such as: PCR-assisted diagnosis, population studies, phylogenetic analysis, and molecular genetics assays.
  
  
  OBJECTIVE
  To evaluate quality and quantity of the P. insidiosum gDNA extracted by three separate protocols intended for fungal gDNA preparation.
  
  
  METHODS
  Seven P. insidiosum isolates were subjected to gDNA extraction by using conventional-extraction, rapid-extraction, and salt-extraction protocols.
  
  
  RESULTS
  The conventional protocol offered the best gDNA in terms of quality and quantity, and could be scaled up. The rapid-extraction protocol had a short turnaround time, but the quality and quantity of the gDNA obtained were limited. The salt-extraction protocol was simple, rapid, and efficient, making it appealing for high throughput preparation of small-scale gDNA samples.
  
  
  CONCLUSIONS
  Compared to rapid-extraction protocol, both conventional-extraction and salt-extraction protocols provided a better quality and quantity of gDNA, suitable for molecular studies of P. insidiosum. In contrast to the other two methods, the salt-extraction protocol does not require the use of hazardous and expensive materials such as phenol, chloroform, or liquid nitrogen.
authors.forename: Tassanee
authors.initials: T
authors.lastname: Lohnoo
authors.forename: Nujarin
authors.initials: N
authors.lastname: Jongruja
authors.forename: Thidarat
authors.initials: T
authors.lastname: Rujirawat
authors.forename: Wanta
authors.initials: W
authors.lastname: Yingyon
authors.forename: Tassanee
authors.initials: T
authors.lastname: Lerksuthirat
authors.forename: Umporn
authors.initials: U
authors.lastname: Nampoon
authors.forename: Yothin
authors.initials: Y
authors.lastname: Kumsang
authors.forename: Pornpit
authors.initials: P
authors.lastname: Onpaew
authors.forename: Piriyaporn
authors.initials: P
authors.lastname: Chongtrakool
authors.forename: Angsana
authors.initials: A
authors.lastname: Keeratijarut
authors.forename: Tristan T
authors.initials: TT
authors.lastname: Brandhorst
authors.forename: Theerapong
authors.initials: T
authors.lastname: Krajaejun
mesh_terms: D000818:Animals; D004247:DNA / Q000302:isolation & purification*; D016678:Genome; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D058968:Pythiosis / Q000235:genetics; D011775:Pythium / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D004247:DNA