Effects of thermal-acid treatment on degradation and amplification of wheat and maize DNA
2018
Bitskinashvili, Kakha | Gabriadze, Inga | Kutateladze, Tamara | Vishnepolsky, Boris | Mikeladze, David | Datukishvili, Nelly
Food processing causes DNA degradation and may hinder polymerase chain reaction (PCR) based detection of food components. Effects of heating (100 °C) and low pH (2 and 4) on degradation and amplification of maize and wheat DNA were investigated. DNA degradation was assessed by agarose gel electrophoresis and PCR amplification. PCR amplicons of different length ranging from 67 bp to 550 bp specific to plant chloroplast, maize or wheat genomes were used to monitor extent of DNA degradation. Combined thermal-acid treatment intensified time-dependent DNA deg- radation observed at thermal treatment and more significantly affected amplification than heating. Increase of acidity from pH 4 to pH 2 had negative influence on the integrity and amplifiability of genomic DNA. PCR amplicons were degraded in a time- and size-dependent manner. Small DNA fragments ranging from 67 bp to 140 bp were amplified in all samples while 226 bp, 259 bp and 550 bp amplicons were not detected in certain highly processed samples. Our findings demonstrate that processing parameters such as temperature, pH and exposure time as well as amplicon size have crucial importance for DNA integrity and accurate detection of food components.
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