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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