Pospisilova Eliska
University of Veterinary and Pharmaceutical Sciences, Czech Republic
Title: comparison of DNA extraction from fresh and processes tuna muscle tissue
Biography
Biography: Pospisilova Eliska
Abstract
Authentication of tuna fish products is necessary to assure consumers of accurate labelling of food products. The most common methods that can be used to identify tuna species include methods based on detection of specific DNA (PCR and its modification: real-time PCR, digital PCR etc.) The quality of species specific DNA crucially affect the efficiency of amplification during the subsequent PCR. The question in DNA detection in processed products lies in the possibility of the DNA fragmentation during the processing technologies and the use of ingredients that may inhibit the PCR reaction. In this study three DNA extraction method were compared: DNeasy Blood and Tissue Kit, DNeasy mericon Food Kit and Chemagic DNA tissue 10 Kit. The quantity and quality of DNA were estimated by measuring DNA concentration and ratios A260/A280. The amplifiability was tested by using a set of primers designed to detect the fragments of COI gene of different sizes ranges from 100 bp to 500 bp. Several parameters were evaluated: the effect of heat treatment (boiling at 70ºC and 90ºC), cold and hot smoking, sterilization procedure of whole and mechanically treated muscle used in canned process (high temperature in combination with high pressure), and pate and spread.