Izvorni znanstveni članak
https://doi.org/10.18047/poljo.21.1.sup.47
COMPARISON OF COMMERCIAL DNA KITS AND TRADITIONAL DNA EXTRACTION PROCEDURE IN PCR DETECTION OF PORK IN DRY/FERMENTED SAUSAGES
Ivona Djurkin Kušec
orcid.org/0000-0002-1363-8447
; Josip Juraj Strossmayer University of Osijek, Faculty of Agriculture in Osijek, Kralja Petra Svačića 1d, 31000 Osijek, Croatia
Žarko Radišić
; Josip Juraj Strossmayer University of Osijek, Faculty of Agriculture in Osijek, Kralja Petra Svačića 1d, 31000 Osijek, Croatia
Miodrag Komlenić
; Food Technol. - Belje Inc, PC Baranjka, Bele Bartoka 33, 31300 Beli Manastir, Croatia
Goran Kušec
; Josip Juraj Strossmayer University of Osijek, Faculty of Agriculture in Osijek, Kralja Petra Svačića 1d, 31000 Osijek, Croatia
Sažetak
In the present study four commercially available DNA extraction kits (Wizard® Genomic DNA Purification Kit, High Pure PCR Template Kit, DNeasy mericon Food and GeneJET PCR Purification Kit), as well as standard phenol/chloroform isolation technique have been evaluated regarding their concentration, purity and suitability for amplification of porcine DNA in dry/fermented sausages. The isolates were assessed for quantity and quality using spectrophotometer (IMPLEN GmbH, Germany). To verify template usability and quality of isolated DNA, the polymerase chain reaction (PCR) targeting at porcine cytochrome b by species specific primers was used. The comparison of extraction methods revealed satisfactory efficiency and purity of all extraction kits, while with standard phenol/chloroform isolation method high concentrations of DNA with low A260/280 were obtained. However, all the investigated techniques proved to be suitable for identification of porcine DNA in dry/fermented sausage. Thus, the standard phenol/chloroform DNA extraction method, as the cost-effective one, can be recommended as a good alternative to more expensive isolation kits when investigating the presence of pork DNA in dry/ fermented meat products.
Ključne riječi
dry/fermented sausage; DNA; extraction method; pig
Hrčak ID:
150920
URI
Datum izdavanja:
2.9.2015.
Posjeta: 7.001 *