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                                                                                                                           Lab 11 
                                                     Bacterial Transformation
1. What are plasmids? How might plasmids be beneficial to bacteria?
2. What occurs during bacterial transformation? How has this process been used to benefit humans? Conversely, how has this process permitted the evolution of "super" bugs that are resistant to multiple types of antibiotics?
3. Describe in detail the plasmid used in your experiment. Be sure to include pGLO, the arabinose operon, and the bla gene. Why does the presence of arabinose influence the expression of the pGlo gene (explain the function of the operon). Why is the bla gene included in the plasmid?
4. Did the plasmid "glow" when exposed to the UV light prior to the transformation experiment? Why or why not?
5. You streaked your transformed bacteria onto four different plates. List them.
6. What was your hypothesis as to whether or not you would achieve bacterial growth on each of these plates? Justify your hypothesis.
7.  (a scenario similar to this one will be asked) Regarding the results of your transformation lab, if bacterial growth occurred on a plate with ampicillin, what can you conclude?
8. If there was a mutation that caused the AraC binding protein to be mutated, how might this affect your results?
9. The plates will "glow" only when the GFP protein is expressed. Can this occur in the absence of arabinose? Why or why not?
10. The plasmid you used included the bla gene because it is used as a selectable marker. What does this mean?
How did you conclude that the bla gene was successfully producing  beta lactamase? 

A couple of quick questions on electrophoresis:
1. The D1S80 locus you amplified via PCR exhibits sequence variability among humans. This fact can be used to identify the source of a DNA sample using Gel Electrophoresis. How? (describe why this electrophoresis process is used but no need to include the details re the process itself)
2. The process applies DNA fragments to a gel and then subjects these fragments to an electrical current. Why?
3. What causes some fragments to move farther up the gel than others?

4. Why is it important to include a DNA "ladder" in your gel?
5. What is the purpose of adding the loading dye to your DNA sample?
6. If you add to your gel DNA from an unknown source along with DNA from several identified sources, what are you looking for after your run the gel? (discuss the comparison of the banding patterns produced by the fragments)   

                               Lab 9 DNA Extraction and PCR Amplification
1. During our DNA extraction protocol, you first added a lysis buffer. What were the contents of this buffer? Why was each of these 3 components required?
2. What was the role of the Proteinase K?
3. Why did you boil your sample after it had time to incubate with the Proteinase K?
4. After extracting your DNA, you amplified (via PCR) a specific locus called _______.
Is this locus a coding region? Why is this locus more likely to accumulate mutations vs a gene locus? The variation in this locus is cause by differences in the number of ______s exhibited at this locus.
6. The PCR mix you added to your extracted DNA contained several components. List them.
7. How was this PCR mix specific to the locus you amplified?
8. Describe the three steps in PCR.
9. After 36 rounds of PCR, one locus of DNA will serve as the template to produce how many copies?  WOW!         
 
                                               Lab 8: Genetics and Blood Typing
                 (for the crosses on the quiz, you'll need to show ALL work to receive credit for an answer)
1. A phenotype is the expression of alleles. What is a genotype?
2. The alleles of the ABO gene display a form of inheritance called _______. A person who is type A could have two possible genotypes, ___ and ____. A person who is blood  type O must be genotype ___ because the O allele is ___ to the A and B alleles.
3. In your lab, agglutination indicated a (positive/negative) reaction. How did you determine (be specific)  if a sample was :
   Type A?
   Type B?
   Type O?
   Type AB?
   How did you determine if a sample was Rh + or Rh -? 
4. The ABO locus (gene), positioned on chromosome 9, and the Rh locus, positioned on chromosome 1, are not "linked". What does this mean? What implications does this have for how these two genes are inherited. Would their inheritance mechanism be different if these two genes were linked? How?  
5. A person who is Rh+ mates with a person who is Rh-. They have an Rh- child. What is the chance that their second child will also be Rh-?
6. A woman who is AB+ has a mother who is type A-. the woman mates with a man who is O-. What is the likelihood they will produce a type A- child?
7. What role to cell surface antigens play in the immune system? Plasma antibodies?
8. Duchennes Muscular Dystrophy is due to a recessive allele of an X-linked gene. A carrier female mates with an unaffected male. What is the likelihood they will produce a female child who is also a carrier?
9. In rabbits, white fur is recessive to brown fur and red eyes are recessive to brown eyes. A rabbit that is heterozygous for fur and homozygous recessive for eye color mates with a rabbit that has white fur and is heterozygous for eye color.  What is the likelihood (percent or fraction) they will produce a bunny that has brown fur and brown eyes?
10.  Contaminating the antiserum samples by repeatedly using the same pipette tip across samples would be a drag. How might this poor lab technique alter your results?


                                                                Lab 7: Photosynthesis
 1. What is the balanced formula for photosynthesis?
 2. The process of photosynthesis is described in two reactions. They are the ___ and ___ reactions. 
 3. What are the reactants and products of each of these reactions?
 4. Light travels in a wave pattern. Wavelength measures the distance between the peaks(or troughs) of a wave. How is wavelength related to the energy that wave carries?
 5. Which approx. wavelength of visible light is utilized least by a plant? By looking at a plant, how can you tell this wavelength can not be used by a plant efficiently during photosynthesis?
 6. The technique we used to separate pigments is called ___. Describe the principle behind this technique (include a discussion of how molecular weight/solubility for each pigment factor into the separation of the pigments.)
 7. You identified four pigments via this technique. List them. 
a.
b.
c.
d.
 8. How did you calculated a pigment's Rf value? What does the Rf value represent? Of those four pigments identified, which pigment has the largest Rf value? the smallest? 
 9. Following the chromatography process, you eluted each pigment to produce a sample that was then analyzed by the spectrophotometer.  What is a spectrophotometer designed to measure? The spec produced a graph for each eluted pigment. The Y axis was the ____ and the X axis the ____. What was the graph designed to illustrate? 
 10. If you obtained a curve for the Y-axis near zero (no peaks), what might have been a plausible answer as to why this occurred?  If you could adjust the Y axis scale to get a better result, what would you do to the scale? 


Lab 7: Statistics
1. What is the difference between a null and alternate hypothesis. Given an example of each.
2. What type of data would you use a T-test to analyze?
3. What is meant by variance? Why might variance be taken into account when comparing two mean values?
4. What test examines the variance between two sample populations?
5. A representative sample is often used instead of a complete sampling when testing a hypothesis. Still, statistical tests must rely on an adequate sample size. Why do you think having an adequate sample size is important when analyzing data?
6. What is meant by the p-value?
7. What maximum p-value suggests the data is statistically significant? Would you accept or reject the null hypothesis at this p-value?
8. Degrees of freedom may be thought of as the number of "observations" in the data that are free to vary when estimating statistical parameters. How do you determine the degrees of freedom in a chi-square test? As the "d of f" increases, the X2 number must ________ to maintain a p-value that indicates the data is statistically significant. (refer to the X2 table)  
9. You will be given a simple data set to analyze for fitness using a simple chi-square test.    


                               Lab 6: Measuring Cellular Respiration via Titration
1. What is the formula for aerobic cellular respiration? What is the formula for anaerobic respiration carried out by yeast?
How does this differ from the process carried out by animals? Does anaerobic or aerobic respiration result in a higher net gain of usable cell energy re ATP?
2. A by-product of cellular respiration is carbon dioxide. When is Co2 produced during cellular respiration? How does this product alter the pH of water? By measuring CO2 production, you can infer the rate of cellular respiration. Why? 
3. What indicator did you use to test for the presence of carbon dioxide? How does t is indicator work? 
4. You used _________ as the neutralizing solution for your titration. Why?
5. The control you used in your titration lab was________ . Why did you use this as a control?
6. How did you determine the volume of the organisms use in your titration experiment? Why as it necessary to do so?
7. The experimental beaker containing the plant was placed in the dark during the measurable period of respiration. Why?
     Does this really matter?
8. What did your data show? What this what you expected?
9. List any extraneous variables that may have altered the quality of your DV?       



                                                           Lab 4: Diffusion/Osmosis 
                                                                                 and
                                                                   Lab 3: Microscopy   
 1. Are you comfortable focusing a slide to high power using the correct  technique? (refer to the "How to Use a 
    Microscope" outline for guidance)
2. Are you able to explain how to calculate total magnification?
3. What is meant by "field of view"?
4. Under which power should you ONLY use the course adjuster knob? Why?
5. What is diffusion? Osmosis?
6. A cell containing a 40% sucrose solution is placed in an environment exhibiting a 70% sucrose solution.
    What happens to the cell? Why? What is the  tonicity of this system?  
7. An animal cell in a hypotonic system may result in the cell bursting. Why?  
8. Plasmolysis occurs when a plant cell is placed in a hypertonic state. What is plasmolysis? 
9. What is meant by the term concentration gradient? How is a system's concentration gradient related to its
     rate of osmosis? How did our class data set illustrate this concept?
                                                                        
                                                
                                                                                        Lab Two:
                                                      Macromolecule Identification
1. You tested an unknown sample of food for the presence of four different nutrient molecules. List them. 
2. Polymers are composed of smaller molecules called monomers. List the monomers that are used to construct the following polymers: proteins, polysaccharides, triglycerides (lipids).
3. Organic polymers are constructed from monomers via a reaction called _________________. This is the opposite of the reaction used to separate polymers into their smaller monomer units. That reaction is called _____________.  What role does water play in these synthetic or catabolic reactions? 
4. What negative control did you use in each of the four nutrient tests? Why was it necessary to use a positive and negative control for each of the nutrient tests? What was the positive control in each of the four tests?
5. What indicators did you use in each of the four tests? Explain why each of these different indicators change color when a certain nutrient molecule is present.   
6. What does pH measure? A low pH (<7) number indicates the solution is __________.

A high pH (>7) number indicates the solution is _________________.  

                                                                                Lab One:
 1. Describe the Scientific Method.
 2. Provide an example of a null hypothesis.
 3. In an experiment, what is the dependent variable? The independent variable?
 4. Why is it necessary to have controls in your experiment?
 5. You have just invented an amazing laundry spot remover. You are confident it is better than any   competitor on the market but must prove this to your marketing team. Design an experiment   evaluating the effectiveness of this laundry spot remover. Identify your hypothesis, your dep/indep   variables, any controls you think must be in place, how you will collect your data, and how you will   present your data so your marketing team will easily understand your results. 
 6. What are the base metric units for weight, volume, temperature and length?
 7. Convert:
 23.54 mg = _________ g
 628.7 l = _________ ml
5.6ml = _________ l
 3.6 cm = _________ m
 2,641 m = _________ mm
 8. Record your answers above in scientific notation.  
 

Goals for our Biology 211 Lab
These goals questions are similar to the questions
you will see on your lab quizzes. Please reference these questions to make the most of your study time.
​Your last assessment will rely on the goals questions from labs 11 (with a few electrophoresis questions) and 9 posted below with one question taken from each of the four previous assessments.