DNA Isolation and Analysis Focused on Microorganisms Important in Food Production

Identification of bacterial DNA consists from several steps: cell lysis, isolation and purification of DNA, precipitation by ethanol, identification of bacterial strain by PCR or other molecular biology methods. Each step must be optimised. Nucleic acids can be isolated from cells using magnetic particles. The molecules of DNA are bound to the surface of magnetic carriers by electrostatic interaction, and then they are eluted into buffer. The aim of the work will be to optimize individual steps of identification of bacterial DNA: cell lysis, DNA isolation, characterization of solid magnetic carriers functionalized by amino groups for nucleic acids isolation. The presence of DNA will be verified using agarose gel electrophoresis and the amount of eluted DNA will be determined spectrophotometrically. The quality of isolated DNA will be proved by their amplification using polymerase chain reaction (PCR). Furthermore, the thesis focuses on the study of secondary structures of nucleic acids – cruciforms structures and quadruplexes. These structures are involved in the regulation of cellular processes and their appearance is associated with cancer development and neurodegenerative diseases. In silico genome analysis was performed on important food industry microorganisms. The microorganisms genomic sequences were obtained from the NCBI (National Center for Biotechnology) database. The Palindrome Analyzer and G4 Hunter software were used for the analysis