Vývoj a validace metod pro detekci GMO

• Main Author: Hodek, Jan
• Other Authors: Ovesná, Jaroslava
• Format: Doctoral Thesis
• Language: Czech
• Published: 2014
• Online Access: http://www.nusl.cz/ntk/nusl-332365

The present work is focused on the issue of genetically modified organisms (GMOs) detection in the plants and derived products. Existing legislation in the European Union regulates monitoring of GMO and lays down rules for their labeling and traceability. The aim was to contribute to solving certain issues of critical points in GMO detection. The first point was DNA extraction. The problem was adressed to the example of DNA extraction from the fruit of papaya and candied papaya. For both food products the suitability of the selected extraction methods was verified and confirmed by amplification of a specific DNA sequence by PCR. The second critical point was related to PCR inhibitors. We observed the effect of real-time PCR inhibition due to the influence of the residual EDTA and the residual amount of heavy metal ions, which were present in laboratory plastic. In both examples, the inhibition affects the PCR efficiency. Another critical point in the GMO detection is use of screening elements. That work was focused on false positive results. Development of new methods for GMO detection was another goal of this work. We have developed the method for garden pea taxon-specific gene quantification by real-time PCR. The method was tested on 13 cultivars of garden pea and several related plants of...