Molecular Tools for Studying Genes and Gene Activity

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Molecular Tools for Studying Genes and Gene
Activity

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5.1 Molecular separation 5.2 Labeled Tracers 5.3
Using Nucleic Acid hybridization 5.4 Mapping and
Quantifying Transcripts 5.5 Measuring
transcription rates in vivo 5.6 Assaying
DNA-protein interactions 5.7 Knockout
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Molecular

• Gel electrophoresis
• Two-Dimensional Gel Electrophoresis
• Ion-Exchange Chromatography
• Gel Filtration Chromatography

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Labeled Tracers

• 1Autoradiography
• a mean of detecting radioactive
  compounds with a photographic emulsion
• 2 phosphorimaging----accurate in quantifying
• 3 liquid scintillation counting----using the
  radioactive emissions from a sample to create
  photons of visible light that a photomultiplier
  tube can detect
• 4 nonradioactive tracers----rivaling the
  sensitivity of the radioactive and more security

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Nonradioactive tracers
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Using nucleic acid hybridization

• To name a restriction endonuclease e.g.
  EcoRI
• Frequency of the occurrence of hexamaeric
  sequence
• 1/4096 (4-6)
• Randomly

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Using nucleic acid hybridization
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Using nucleic acid hybridization

• 1 Southern blots identifying specific DNA
  fragment -----Estimating the number of gene family

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Southern blots
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Southern blots
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Using nucleic acid hybridizationDNA
fingerprinting and DNA typing

• Minisatellite DNA----fingerprinting DNA

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Using nucleic acid hybridizationforensic uses of
DNA fingerprinting and DNA typing

• 1 restriction fragment length polymorphism

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Using nucleic acid hybridization

• DNA sequencing----the sanger chain-termination
  sequencing method

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Using nucleic acid hybridization

• Restriction mapping

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Assaying DNA-protein interactions

• Filter binding

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Assaying DNA-protein interactions

• Gel mobility shift
• a small DNA has a much higher mobility in
  gel electrophoresis than the same DNA does when
  it is bound to a protein

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Assaying DNA-protein interactions DNSase
footprinting
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Assaying DNA-protein interactionsDMS
footprinting and other footprinting methods
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Knockouts
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Knockouts