1) The history of gene cloning. Overcoming barriers to the free transfer of genes between organisms. Genetic manipulation. The structure of the genome of prokaryotes and eukaryotes. E. coli in genetic engineering - strains, genotype, genes. 2) Isolation and manipulation of DNA - the isolation of genomic and plasmid DNA, separation and amplification of plasmids. 3) Modification of DNA ends - restriction endonucleases and other enzymes in genetic engineering - types, classification, properties, and buffers. 4) Methods of DNA transfer into the living organisms - transformations, selection, and identification of the transgenes. 5) Cloning vectors - the development, use, types, and differences between vectors for different organisms (yeast, plants, animals). 6) From DNA to construct - How to proceed in isolation of the gene? Genomic library, cDNA library, classical vs. Gateway cloning technology, oligonucleotides. 7) PCR - history, principles, polymerases and their properties, troubleshooting. Sequencing. 8) Cloning and basic research I Study of gene position. Study of gene expression. 9) Cloning and Basic Research II Analysis of gene expression: Northern blot, a brief introduction to microarrays and their use. Site-directed mutagenesis. 10) Cloning of the genes and Basic Research II Reporter genes, RNA interference, protein-protein interactions. 11) Gene therapy. 12) Epigenetics, the final discussion about the new knowledge.
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