Předmět Methods in Molecular Biology (KMB / 770)

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Materiál	Typ	Datum	Počet stažení

Obsah

Contents of lectures:Theoretical lectures will explain principles of the metods and their application. The following topics will be covered:1.Recombinant DNA, cloning vectors, gene libraries.2.Purification and labeling of nucleic acids, hybridization.3.DNA modifying enzymes, cloning strategies, DNA sequencing.4.PCR applications.5.Analyses of mRNA, S1 mapping and primer extension.6.Analyses of gene expression at the mRNA level: in situ hybridization, RT-PCR, quantitative RT-PCR, microarray technology.7.Extraction and separation of proteins.8.Design and preparation of recombinant proteins in E. coli, fusion and tagged proteins.9.Antibodies, immunodetection methods.10.Analyses of gene expression at the protein level: western blotting, immunocytochemistry.11.Protein-protein interaction analyses: GST pull-down assay, immunoprecipitation, yeast two-hybrid system.12.Protein-DNA interaction analyses: EMSA, DNase protection assay.13.Analyses of transcriptional regulation in cell transfection systems.14.Analyses of gene expression and function in transgenic eukaryotic models (Drosophila andC. elegans).Contents of laboratories:Of about 15-20 offered methods that cover most of the lecture topics, each student will choose 3-4. Students will perform these methods in some of the laboratories of the Academy institutes under the supervision of the laboratory personnel. Each method takes between 1-4 days.

Literatura

Alberts et al.: Molecular Biology of the Cell. Sambrook, J., Fritsch, E. J. and Maniatis, T. (1989) Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press.

Požadavky

Periodicita: v každém akademickém roce.Speciální kurz doktorského studijního oboru Fyziologie a vývojová biologie a Molekulární a buněčná biologie a genetika

Garant

Mgr. David Doležel, Ph.D.