Science and Technology Production
Protocolo de Extracción de ADN Optimizado para cepas de Staphylococcus aureus Utilizando Nitrógeno Líquido Optimized DNA Extraction Protocol for Staphylococcus aureus Strains Utilizing Liquid Nitrogen
Articulo
Authorship:
Galeano Mariana Belen ; Robaldi Stefania ; Gordillo Tania ; Ricardi Martiniano M ; Cassanelli Pablo M ; Pereda Rosana ; Palomino Maria Mercedes ; TRIBELLI, PAULA MARIADate:
2025Publishing House and Editing Place:
ASOCIACION ARGENTINA MICROBIOLOGIAMagazine:
REVISTA ARGENTINA DE MICROBIOLOGÃA ASOCIACION ARGENTINA MICROBIOLOGIASummary *
DNA extraction is crucial for conducting experiments such as whole-genome sequencing and demands methods that are reproducible and cost-effective. Lysing Staphylococcus aureus cells is particularly challenging due to its peptidoglycan layer resistant to common treatments. Traditional methods involve costly enzymatic lysis with lysostaphin. Here, we developed a novel approach utilizing liquid nitrogen and mechanical disruption in a mortar for lysis. DNA from S. aureus USA300 and clinical isolates was purified using phenol-chloroform extraction followed by precipitation. The integrity and purity of the DNA were confirmed, obtaining suitable concentration and purity for various molecular biology techniques. The quality of the employed DNA was validated by amplifying fragments of different genes using PCR. This method circumvents lysostaphin, yielding DNA suitable for use in other techniques. Information provided by the agent in SIGEVAKey Words
ADN EXTRACTIONSTAPHYLOCOCCUS AUREUS