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Part I.3 Methods in Molecular Cell Biology

Part I.3 Methods in Molecular Cell Biology. 304515 Molecular Biology of the Cell Teacher: Montarop Yamabhai http://www.sut.ac.th/AgriTech/Biotech/mywebpage.html. Manipulating cells in the culture. Isolating cells Disrupt extracellular matrix with proteolytic enzymes or EDTA

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Part I.3 Methods in Molecular Cell Biology

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  1. Part I.3Methods in Molecular Cell Biology 304515 Molecular Biology of the Cell Teacher: Montarop Yamabhai http://www.sut.ac.th/AgriTech/Biotech/mywebpage.html

  2. Manipulating cells in the culture • Isolating cells • Disrupt extracellular matrix with proteolytic enzymes or EDTA • Fluorescence-activated cell sorter • Growing cells in culture dishes • Primary cell culture • Prepare directly from tissues of an organism • Cell line • Immortalized, can grow indefinitely

  3. Fractionation of Cells • Organelles and macromolecules can be separated by ultracentrifugation • There are 2 types of sedimentations • Velocity sedimentation • Equilibrium sedimentation

  4. DNA Engineeting

  5. Analyzing Protein Structure and Function • Chromatography / HPLC / FPLC • Ion-exchange/hydrophobic chromatography • Gel-filtration chromatography • Affinity chromatography • Gel Electrophoresis • Western Blotting • Mass spectrometry • X-ray crystallography/NMR spectroscopy • Protein-protein interaction • Protein-DNA interaction

  6. Determination of size and subunit of a protein by SDS polyacrylamide-Gel Electrophoresis

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