The effect of different rapid methods for isolation of Salmonella genomic DNA on PCR sensitivity | Vpliv različnih hitrih metod osamitve genomske dnk salmonel na občutljivost PCR
2001
TRKOV, Marija | JERŠEK, Barbara
English. The purpose of our research was to determine the effect of rapid methods for isolation of Salmonella genomic DNA on PCR sensitivity. Salmonella specific primers ST11 and ST15 were used for DNA amplification. Among tested procedures the best PCR sensitivity was achieved after treating of sample with NaOH and SDS solutions and heating of sample in water bath at 95o C for 15 min. PCR detection limit for Salmonella in pure and mixed cultures was in a range from 104 to 105 cfu ml-1. Described procedure for DNA isolation is rapid and not work-consuming.
Show more [+] Less [-]Slovenian. Namen raziskave je bil proučiti hitre metode osamitve genomske DNK salmonel in določiti njihov vpliv na občutljivost PCR. Za pomnoževanje DNK sta bila uporabljena za rod Salmonella specifična začetna oligonukleotida ST11 in ST15. Izmed preskušenih postopkov je bila najboljša občutljivost PCR v primeru, ko smo spranim celicam dodali mešanico raztopin NaOH in SDS ter vzorec 15 minut segrevali v vodni kopeli pri 95o C. Prag občutljivosti PCR za določanje salmonel tako v čisti kot v mešani kulturi je bil v območju 104-105 cfu ml-1. Opisan postopek osamitve DNK je hiter in izvedbeno ni zahteven.
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