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  1. 20th century was a century of __________ in Biology.
  2. 21st century is a century of ________ in Biology.
  3. Describe Neural Circuits.
    • They underlie all aspects of behavior.
    • A wide variety of structures and regulatory proteins are required for a single circuit.
    • Simple organisms have simple neural circuits and vice versa.
    • Even subtle changes in expression of a single gene can affect behavior.
  4. Subtle changes in the expression of a single gene can affect behavior, such as:
    • Absence of expression
    • overexpression
    • changes in degree of expression
    • expression of splice isoforms
    • changes in expression pattern
    • can overlap and take care of several functions
  5. What is Phenylketonuria? What's it an example of?
    • It is the inability to break down the AA phenylalaine.
    • It is a recessive autosomal inherited trait.
    • It's an example of a diseas caused by the change of a single gene.
  6. What are some model organisms in Neural Science?
    Mice, Elegans, Fruit flies, monkeys, frogs.
  7. Journals that publish neuroscience:
    • Journal of Neuroscience
    • Nature Neuroscience
    • Behavioral Neuroscience
  8. Where is genetic information stored?
  9. How many chromosomes do humans have? What kind?
    46. 22 pairs of autosomes and a 2 sex chromosomes.
  10. What do genes do?
    • Regulate behavior
    • Emotional regulation of behavior
  11. Human brain and _____ brain react ______ to ________ stimuli in the ______ (region of the brain).
    Mice ; Similar ; fear ; amygdala
  12. Two major genetic approaches for studying specific DNA sequences
    • 1. Specific amplification (DNA cloning)
    • 2. Specific detection
  13. Cloning of PCR products. (Types)
    • Blunt-end cloning
    • TA cloning (A- over hang)
    • incorporation of restriction sites in PCR oligoprimers
  14. Classical methods to detect and analyze gene transcription products
    • Northern blotting
    • rt-PCR/Q-PCR
    • cDNA library
    • RNA in situ hybridization
  15. Northern Blotting
    • + Quantitative. Size. Integrity. Amount
    • -Not sensitive
  16. RT-PCR/Q-PCR
    • Beneficial because you can isolate small amounts of RNA.
    • +Quantitative. Very Sensitive.
    • -Need to know sequence by designing primers.
  17. pros and cons of RNA in situ hybridization
    • +Allows to analyze where a gene of interest is expressed in the brain. Can be semi-quantitive if radioactive.
    • -Not always possible.
  18. "Sense"
    • Makes proteins.
    • Use antisense probe.
  19. "Antisense"
    • Doesn't make protein.
    • Use sense probe.
  20. Steps of RNA in situ hybridization
    • 1. Cut brain sections and place on glass slides using cryostat at -25oC
    • 2.DNA or RNA labeling. DNA-oligo, Use 33P or 35S. RNA- Use dioxigenin-UTP. 33P or 35S.
    • 3.Put on the brain slice and let it hybridize over night at 60oC. Then wash away probe.
    • 4. Observe with radioactive x-ray film.
  21. Why can't 32P be used for in situ?
    • It is a very high energy isotope, too much for in situ.
    • Can use for Northern and Southern blotting.
  22. Radioactive oligo in situ hybridization
    • DNA is labeled using 33P or 35S radioactive dNTP transferase enzyme.
    • Oligo is applied to the brain sections in a hybridized buffer over night.
  23. NeuN is a ___________ in ____ neurons.
    • protein ; all.
    • Serves as a marker.
  24. Visualization of gene expression in vitro and in vivo uses
    • LacZ
    • GFP (green fluorescent protein)
  25. start at KO mice "methods to create a genetically modified animal..."
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