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Chapter 21: Nucleic Acids: Isolation, purification, detection, and Hybridization Purification of DNA 21.01: Phenol extraction removes proteins form nucleic acid Removal of unwanted RNA 21.02: Removal of RNA by Ribonuclease Gel electrophoresis of DNA 21.03: Principle of electrophoresis 21.04: Agarose gel electrophoresis of DNA 21.05: Agarose gel electrophoresis of DNA: staining and standards 21.06: Pulsed field gel electrophoresis (PFGE) Denaturing Gradient Gel electrophoresis (DGGE) 21.08: Parallel and Perpendicular DGGE 21.09: Chemical synthesis of DNA on glass beads-Priniciple 21.10: DNA synthesizer 21.11: Phosphoramidite nucleotides are used for chemical synthesis of DNA 21.12: Addition of spacer molecule and first base to glass bead 21.13: Chemical synthesis of DNA-Nucleotide addition 21.14: Chemical synthesis of DNA-Coupling Chemical synthesis of complete genes 21.15: Synthesis and Assembly of a Gene Peptide Nucleic Acid 21.16: Structure of peptide nucleic acid compared to DNA Peptide Nucleic Acid 21.17: Triple helix of peptide nucleic acid with DNA Measuring the concentration of DNA and RNA with UV light 21.18: Absorption of UV radiation by nucleic acids Radioactive labeling of nucleic acids 21.19: Use of 32P and 35S to label nucleic acids Detection of radio-labeled DNA 21.20: Scintillation counter is used to measure radioactivity 21.21: Autoradiography to detect radio-labeled DNA or RNA Fluorescence in the detection of DNA and RNA 21.22: Fluorescence detection 21.23: FACS machine can sort chromosome Chemical tagging with biotin or digoxigenin 21.24: Labeling DNA with Biotin 21.25: Detection systems for biotin The electron microscope 21.26: Principle of the electron microscope 21.27: Metal shadowed DNA molecule are visible under an EM 21.28: R-loop analysis to visualize introns Hybridization of DNA and RNA 21.29: Melting of DNA at high temperature 21.30: Hybrid DNA formed by melting and re-annealing 21.31: Relatedness of DNA by filter hybridization Southern, Northern and Western Blotting 21.32: Southern blotting: DNA+DNA hybridization Zoo Blotting 21.33: Zoo blotting reveals coding DNA Fluorescence in Situ hybridization (FISH) 21.34: FISH-Principle 21.35: FISH to localize genes on chromosomes 21.35: FISH to detect and measure mRNA levels 21.37: Molecular Beacon