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44 Cards in this Set
- Front
- Back
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What is the name of the liquid that turns into a solid gel?
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liquid agarose
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What is the gel tray submerged into?
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TBE (gel running buffer)
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What were the four types of samples that we tested DNA on?
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Standard, Unknown, Onion, and Lambda DNA
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What voltage is the electrophesis metere set to?
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100 volts
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Determining the size and density of DNA fragments is known as ... analysis
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quantitative
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Does DNA have a negative or positive charge?
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negative
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What are the three enzymes we used to "cut" the Lambda DNA?
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EcoRI, Hind III and Bgl II
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How can you figure out the size of unknown samples? (hint: graph)
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standard curve graph using fragments of already known size
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What is inverse relationship?
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Fragments that are shorter will migrate farther through the gel
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On a standard curve, what is plotted on the y axis? The x axis?
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y=fragment size (bp)
x=migration distance (mm) |
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When measuring migration distance, you measure from what to what?
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well to fragment
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DNA digested by Hind III produced how many fragments?
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9
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What is the speacil graph paper that we used called?
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cycle semilog paper
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Generally speaking, on our graph, as the migration distance got larger, what happened to the fragment sizes?
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they got smaller
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What is the term we use when "something" in one cell causes change in another cell? What is this "something" called?
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transformation
dna |
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Transformation can "force" another cell to express a protien it would not normally produce. T or F?
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true
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What is the bacteria that is commonly used in transformation?
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E.coli
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What temp does E.coli grow at?
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37C
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What is commonly used to "introduce" a gene of interest into another cell?
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plasmids
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What are plasmids?
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small circular fragments of DNA that are able to replicate on their own
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What do antibiotics do? What exactly are antibiotics?
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kill bacteria
fungi toxins |
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What allows bacteria to be resistant to antibiotics?
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plasmids (genes)
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When bacteria resists antibiotics, what is actually happening?
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it is resisting it's cell wall be "broken" by the antibiotic
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Bacterial cells that have the "resistant" gene are known as what?
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drug resistant
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Drug resistant bacteria are identified by their presence of plasmids. Therefore, this is a ... marker.
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selectable
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What is ampicillin? What does it do?
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1. an antibiotic
2. it inhibits the production of the cell wall on bacteria (therefore it is letha. to bacteria) |
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Is it possible for ampicillin to NOT BE leathal to bacteria? Why?
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yes
bacteria can contain the plasmid which makes it resistant |
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How are plasmids typically abbreiviated?
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p (followed by some info about the gene being introduced)
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In our lab, what is the name of the plasmid that allows bacteria to be resistant to ampicillin?
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pAMP
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What is pGLO? What gene does it contain?
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1. it is a plasmid that can be placed into bacteria
2. it contains the gfp gene which produces a protein that makes the bacteria "glow" |
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The pGLO gene (gfp) was inserted into what plasmid in our lab?
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pAMP
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The gfp gene is normally "off". What will turn it on?
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if it is in the presence of sugar (arabinose)
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The sugar arabinose activates what gene?
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araC gene
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If bacteria is put on a plate containing the sugar arabinose AND the antibiotic ampicilin - what will happen to the bacteria that have the pGLO plasmid? Why?
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1. it wil grow AND glow
2. because of the pGLO plasmid (which contains pAMP) it will grow - it makes it unaffected by the antibiotic. Because the sugar activates the gfp gene in pGLO, it will glow! |
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What is the pathway that activated the glow?
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Must have sugar (arabinose) which activates the gene araC. This in turn activates the gfp gene which creates the glow.
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Bacterial cells must first be made ... in order to insert plasmids into them.
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competent
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Is DNA hydrophillic or hydrophobic?
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hydrophillic (water loving)
therefore it does not easily cross the bacterial cells membrane |
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How do we more easily force DNA through the bacterial cell membrane?
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Through calcium chloride heat shock which neutralizes the DNA's negative charge
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When using the 1000 ul micropipeter, how would you gauge 730 ul? 1000ul?
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0 1
7 0 3 0 In thousands, the last number is dropped |
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The LB agar plate ONLY with plasmids will do what? Why?
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1. it will grow ONLY
2. the plasmid contains the pAMP which is makes it resistant to the antibiotic. it will NOT glow because there is no sugar (arabinose) to activate the gfp gene |
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The LB agar plate with plasmids, AMP and arabinose will do what? Why?
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1. It will glow AND grow
2. It will grow because the bacteria contains the plasmids that make it resistant to AMP. it will glow because it contains arabinose (sugar) which activates araC gene which in turn activates the gfp gene causing it to glow |
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The LB agar plate with no plasmids and no pAMP will do what? Why?
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1. It will grow but it will not glow
2. it will grow because no AMP was added - AMP is what inhibits the formation of the bacterial cell walls. it will NOT glow because there are no plasmids with pGLO |
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The LB agar plate with AMP and arabionse (sugar) but NO plasmids will do what?
Whay? |
1. It will neither grow nor glow
2. it will not grow because AMP will inhibit the growth of the bacterial cell wall and there are no plasmids to resist this. it will no glow because there are no plasmids with pGLO. The arabinose is therefore useless because it has nothing to "activate". |
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What will be doing in today's lab?
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1. Recording the colonies
2. Identifiying which have grown and which ones glow 3. Will be adding the GLOWING bacteria to a test tube containing AMP and another test tube containing AMP and arabinose 4. Will calculate the mass of plasmid DNA and the number of colonies transformed per microgram of plasmid DNA |
