dec 1 DNA hybridization - Enzyme P 32 P - dATP Direct...

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Nucleic Acid Hybridization 12/1/06 Major ideas: 1. The hydrogen bonds in nucleic acids can be broken and reformed. These processes are called denaturation and renaturation or hybridization . 2. Using your knowledge of nucleic acid structure and properties, you should be able to determine whether specific single-stranded nucleic acids, DNA or RNA, can hybridize to each other. 3. You should be able to determine the kinds of information that can be obtained from DNA hybridization and from Northern and Southern blots
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1 Hybrid Detection of Hybrids Label the probe with a detectable marker: Radioactive Fluorescent
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Unformatted text preview: Enzyme P 32 P - dATP Direct labeling Fluorescent dye labeled Biotinylated dUTP Indirect Labeling Streptavidin binds biotin HRP conjugated to streptavidin can be detected with a substrate 2 How is the label incorporated into the DNA? Helicase Fig. 9.5a1 Fig. 9.5a2 mRNA can also be separated by gel electrophoresis 3 Southern (DNA) or Northern (RNA) blotting for detection of specic fragments. Fig. 9.15d DNA bound to nitrocellulose Add labeled probe Northern blot example showing SRY candidate for testes determining factor is expressed in testes, but not lung, ovary, or kidney...
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dec 1 DNA hybridization - Enzyme P 32 P - dATP Direct...

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