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The influence of polysaccharide contamination on molecular-biology analysis
Žylková, Kateřina ; Strečanská, Paulína (referee) ; Fialová, Lenka (advisor)
The presence of polysaccharides in DNA isolation and its subsequent analysis often leads to undesirable effects. Polysaccharides together with other metabolites (phenolics, proteins) can act as inhibitors of PCR. In this work, the effect of polysaccharide contamination on the analyzed DNA was investigated. In the experimental part, DNA samples were isolated from two exotic fruits (mango, banana), from which the concentration of polysaccharides was then determined. The analysis showed that by adding CaCl2, the polysaccharide content of the samples was significantly lower. After checking the amplification of the DNA samples with added CaCl2, it was found that CaCl2 itself inhibited PCR and therefore had to be removed from the samples. After purification, the amplification of the DNA was reverified and it was found that the DNA with CaCl2 after purification gave the best results. These results were further verified by agarose gel electrophoresis, which confirmed that a reduction in the polysaccharide content of the samples helped DNA amplification. It was also observed that it depends on the type of polysaccharides present in the source plant material. Banana DNA showed better amplification results than mango DNA. This is due to the different chemical composition of these fruits. Banana, unlike mango, does not contain polysaccharides that would significantly contaminate the isolated DNA.
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