Labs 1-12

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Labs 1-12
2011-04-25 17:52:44
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  1. For zucchini cores, what effect does adding more solute do?q
    it decreases water potential around the core causing more water to diffuse out of it. This will result in a higher % decrease in mass.
  2. What does the dark bottle represent?
  3. What does the initial bottle in Lab 12: DO represent?
    amount of DO there is to begin with
  4. What is NPP?
    • GPP- R or Light- Initial
    • o amount of organic compounds that is available to the animal after photosynthesis and respiration has occurred; change over time in DO from initial concentration
  5. Bacteria can only take up DNA during the period at the end of logarithmic growth called __.
  6. __ can transfer genes that occur naturally within them, or __ can act as __ for introducing foreign DNA from other bacteria, plasmids, or even eukaryotes.
    plasmids x2
  7. Uptake of plasmids is a way of bacteria expressing foreign DNA. In a sentence explain what physical process must be done to the bacteria to make it compentent.
    • you can incubate them
    • growing more rapidly causes them to become more competent
    • -- heat shock in presence of .05mg of CaCl2. You place the CaCl2 in, then you put the bacteria in a heat bath at 42oC- 45oC, wait a couple of minutes and immediately transfer then to an ice bath. THis is the heat shock and makes their membranes more receptive to take up plansmids.
  8. What does competent mean?
    making a membrane more receptive and vulnerable so that it can take up a plasmid; when a cell takes up fragments of DNA that are attached to its membrane
  9. Explain the process that occurred that allowed E. Coli to become ampecillin resistant.
    • bacterial transformation
    • - made competent
    • - competent cells were placed in two vials: one with the plasmid and one without
    • - we allow the experiment to go, allowing the cells to take up the plasmid; not all cells take up the plasmid
    • - afterwards, we have four petri dishes; two with ampicillin and two without ampicillin
    • - we take the cells in the ampR+ vial and spread them on the plasmid with ampiciliin and without ampicillin
    • o results: the dish without ampicillin should have a lawn of growth/ the dish with ampicillin should have colonies showing those that took up the plasmid
    • - take the ampR- cells and spread them on the dishes that are labeled no plasmid with ampicillin and without.
    • o Results: the dish without ampicilllin should also have a lawn of growth/ the dish with the antibiotic should have no growth because none of those cells were exposed to the plasmid so they are not ampicillin resistant
    • o Those that took up the plasmid will survive; those that did not will die.
    • o The descendents will have that gene
  10. In Lab 6, did transformation occur? How do you know?
    yes because there were colonies that grew in the jar with ampicillin
  11. WHat other steps can be taken to verify that transformation has occurred?
    other steps can be speading the cells on another jar with the antibiotic; if they still grow, they are resistent
  12. If you had to identify two unknown DNA samples, what steps would you have to do?
    • a. Restriction Enzyme digest - used to cut DNA
    • b. Gel electrophoresis - used to separate different size pieces of DNA
    • c. Denaturing into single-stranded DNA - process used to view only one strand of the DNA double helix
    • d. Southern Blot - DNA bands transferred to a nitrocellulose paper
    • e. Radioactive DNA probe - used to find & bind to the complementary sequence in one or more RLFP's
    • f. Autoradiograph - used to show similarities in DNA samples

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