This work was focused on DNA isolation from cheeses for the use in polymerase chain reaction. First, there was optimised the procedure of homogenisation of different types of cheeses from commercial sources, cell lysis and DNA isolation. DNA was isolated using magnetic microspheres and phenol extraction. It was shown that the DNA was amplified in PCR for domain Bacteria after dilution. Next, there was optimised the procedure of DNA isolation from fresh cheeses and from contaminated fresh cheeses and their pickles. DNA from all samples was amplified in PCR. The presence of DNA of domain Bacteria and yeast DNA was demonstrated. In the last part of the work, there were optimised the preparation of PCR mixtures and bacterial DNA amplification in PCR with primers with clamp (F357-GC and R518). Synthetized PCR products were analysed using DGGE. It was shown that amplicons of DNA isolated from cheeses and pickles differ in positions and numbers. Larger number of bands of different intensities was detected after amplification of DNA isolated from contaminated pickles.
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:216973 |
| Date | January 2013 |
| Creators | Mohelský, Tomáš |
| Contributors | Rittich, Bohuslav, Španová, Alena |
| Publisher | Vysoké učení technické v Brně. Fakulta chemická |
| Source Sets | Czech ETDs |
| Language | Czech |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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