MCB 301 Spring 2015
Experiment 7: Enzymes and Adaptation Lab Notebook Assignment
24 points total
Purpose/Relevance (2 points)
Data/Results (4 points)
 Completed tables 1-4
Discussion (15 points)
 All results are clearly interpreted, hypotheses are made and evaluated for the obtained
results and their deviation from the “expected” results. There is a good synthesis of the
experiment and the results, particularly in the scope of a larger context. (10 points)
 What is tryptone? Why is it important and where is it specifically used with regards to
the experiment you performed? (3 points)
 Some bacteria require certain growth factor(s) (e.g., purines, pyrimidines, amino
acids, vitamins) to be added to the medium in order to grow. Other bacteria can
synthesize the factor(s) from another carbon source, so they do not need the preformed
factor(s) added to the medium. What two ingredients in some of the media used in the
experiment supply vitamins for bacterial growth? (2 points)
Grammar and Format (3 points)
Experiment 8: Lactic Acid Bacteria Lab Notebook Assignment
MCB 301 Spring 2015
65 points total
Purpose/Relevance (2 points)
Data/Results (20 points)
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Observation of any bubbles present in enrichment culture
Measured pH of enrichment
Shapes and arrangements of cells from enrichment culture
Observation of colony appearance on TJA plate
Cell characteristics from TJA plate: Gram reaction; shape and cell arrangement from wet
mount; catalase reaction
Observation of colony appearance on HIA-glucose plate
Cell characteristics from HIA-glucose plate: Gram reaction; shape and cell arrangement
from wet mount; catalase reaction
Observation of colony appearance on HIA-sucrose/NaN3 plate
Cell characteristics from HIA-sucrose/NaN3 plate: cell shape and arrangement
Discussion (40 points)
 All results are clearly interpreted, hypotheses are made and evaluated for the obtained
results and their deviation from the “expected” results. There is a good synthesis of the
experiment and the results, particularly in the scope of a larger context. (30 points)
 Explain the purpose of each of the following in the lactic acid bacteria enrichment (4
points):
 Cabbage
 Glucose
 Salt
 Not screwing the cap on the bottle tightly
 Is the HIA-sucrose-NaN3 medium a selective medium, a differential medium, both
selective and differential, or neither? Explain your answer (include what selective and
differential media are, what organisms are selected/differentiated in our experiment and
how?). (6 points)
Grammar and Format (3 points)
MCB 301 Spring 2015
Experiment 9: Environmental Effects Lab Notebook Assignment
60 points total
Purpose/Relevance (2 points)
Data/Results (14 points)
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Completed Tables 1-5
Discussion (41 points)
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All results are clearly interpreted, hypotheses are made and evaluated for the obtained
results and their deviation from the “expected” results. There is a good synthesis of the
experiment and the results, particularly in the scope of a larger context. (23 points)
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The discussion should also include whether or not the data you collected as a section
was sufficient for you to make sound conclusions about each organisms tolerance for
temperature, growth temperature, pH and oxygen use. (8 points)
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Summarize the major classifications of organisms for temperature class, pH and oxygen
utilization. (6 points)
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Discuss how the results of the “Effects of Growth Temperature” experiment differs from
the results of the “Effects of Temperature Tolerance” experiment. (4 points)
Grammar and Format (3 points)
MCB 301 Spring 2015
Experiment 10: Sporeforming Bacteria Lab Notebook Assignment
63 points total
Purpose/Relevance (2 points)
Data/Results (15 points)
Anaerobic plates:
Description of growth
Catalase test results, if performed
Aerobic plates (Period 1):
Description of different colonies
Presence or absence of spores in wet mount
Catalase test results
NA plates (Period 2):
Presence or absence of spores in wet mount
Gram reaction
Catalase test results
Post enrichment:
Calculation of amount of glucose needed for cultures
Location of growth in each of the 3 anaerobic tubes (no shaking)
OD of 5 samples
pH of 5 samples
VP test on 5 samples
Discussion (43 points)
 All results are clearly interpreted, hypotheses are made and evaluated for the obtained
results and their deviation from the “expected” results. There is a good synthesis of the
experiment and the results, particularly in the scope of a larger context. (30 points)
 What ingredients are in Nutrient Agar medium (besides agar) and what is the function
of those ingredients? Is the NA medium a selective medium, a differential medium,
both selective and differential, or neither? Explain your answer. (5 points)
 Describe how the catalase test works. Explain how the results of the catalase test is
used in this Experiment. (6 points)
 Describe two ways that the microscope plays an important role in obtaining a pure
culture of Bacillus or Clostridium from an enrichment. (2 points)
Grammar and Format (3 points)
MCB 301 Spring 2015
Experiment 12: Transposon Mutagenesis Lab Notebook Assignment
62 points total
Purpose/Relevance (2 points)
Data/Results (17 points)
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State the total number of colonies obtained on each of the 8 transposant plates (LBKan)
State the total number of potential mutants that you saw (whites, pinks or
combinations).
State the total number of colonies on the LB plate you counted.
Calculate the number of cells/ml of transposants. Show your work.
Calculate the number of cells/ml of recipients. Show your work.
Calculate the transposition frequency. Show your work.
State the number of potential mutant colonies that remained white, pink or a
combination thereof after you re-streaked them. If you didn’t have any of your own
potential mutants and re-streaked using colonies from someone else's plate, be sure
to state this.
Give the Gram stain results of all colonies that were tested.
State all biochemical tests performed and the results of those tests, including all of
the controls.
Discussion (40 points)
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All results are clearly interpreted, hypotheses are made and evaluated for the
obtained results and their deviation from the “expected” results. There is a good
synthesis of the experiment and the results, particularly in the scope of a larger
context. (30 points)
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How does kanamycin work? (2 points)
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Describe how pRL27 confers resistance to kanamycin to its host? (2 points)
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Where is diaminopimelic acid normally found? Be specific. (2 points)
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If a bacterial cell cannot make diaminopimelic acid and it is not available in its
environment, then the cell may die. How? (2 points)
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A bacterial cell contains pRL27, but the cell does not contain the pir gene. Describe
what happens if you produce a culture of the bacteria by growing it LB broth. (2
points)
Grammar and Format (3 points)
MCB 301 Spring 2015
Experiment 13: Biochemical Tests Lab Notebook
43 points total
Purpose/Relevance (2 points)
Data/Results (8 points)
 Known Organism
 Table of each test performed, with result and appearance
 Table of all 12 organisms with a written/illustrated description of each
Discussion (30 points)
 All results are clearly interpreted, hypotheses are made and evaluated for the obtained
results and their deviation from the “expected” results. There is a good synthesis of the
experiment and the results, particularly in the scope of a larger context. (26 points)
 Why is a color change indicating acid production in a fermentation culture more
meaningful than no color change in the same medium? (2 points)
 Explain why a pH indicator that has a lower pH range is not as effective as a pH
indicator with a higher pH range, when using in a test medium that is buffered well. (2
points)
Grammar and Format (3 points)
MCB 301 Spring 2015
Exercise 14: Identification Lab Notebook
47 points total
Purpose/Relevance (2 points)
Data/Results (13 points)
 Isolation of bacteria, both Gram-positive and Gram-negative. TA must initial your lab
notebook in order to receive credit.
 Record your observations of any growth on MacConkey agar
 Gram reaction and shape of both isolates
 Dichotomous key, Be sure to include both organisms (Gram-positive and Gramnegative) in your key. Note that the key must be initialed by the TA before collecting
any biochemical test media.
 Table of biochemical test results for each isolate
 Identification of each of your unknown organisms
Discussion (29 points)
 All results are clearly interpreted, hypotheses are made and evaluated for the obtained
results and their deviation from the “expected” results. There is a good synthesis of the
experiment and the results, particularly in the scope of a larger context. (23 points)
 What is the key step in any successful identification of a bacterium from a natural
source, such as a human with an infection, and why? (2 points)
 Why are morphological characteristics alone not sufficient to identify an unknown
bacterium? (2 points)
 How does knowing the source of a bacterial specimen make its identification easier? (2
point)
Grammar and Format (3 points)
MCB 301 Spring 2015
Experiment 15: Southern Blot Lab Notebook Assignment
65 points total
Purpose/Relevance (2 points)
Data/Results (20 points)
 Labeled photograph of gel
 Final Southern blot membrane
 Make a table of the bands on your photograph showing:
 The size in kilobases
 The DNA content of each band. Is it vector DNA, insert DNA or a combination of
both vector and insert DNA?
 Is the band present on your blot, yes or no?
 Using your data draw a restriction map showing where the sites are located with
respect to one another on the pGEX/rpoD plasmid. Use the map of pGEX as a
reference. Be sure to include the relative location of each restriction site by
specifying the position number or approximate distances in kilobases between all the
sites that you have indicated. If any sites cannot be accurately placed on your map,
explain the problem.
Discussion (40 points)
 All results are clearly interpreted, hypotheses are made and evaluated for the obtained
results and their deviation from the “expected” results. There is a good synthesis of the
experiment and the results, particularly in the scope of a larger context. (30 points)
 Based on the pattern of bands detected on your Southern blot, what is the identity of
your probe? Does your probe contain the rpoD gene sequence? (2 points)
 How do the results obtained using the other probe compare with your results? Are
different bands detected? How does this illustrate the principle of probe specificity? (2
points)
 Describe the plasmid used in this Experiment (e.g., size, genes, etc). (3 points)
 Based on your agarose gel results, were there multiple bands in the lane containing
undigested plasmid DNA? What would multiple bands represent and why would they
migrate differently? (3 points)
Grammar and Format (3 points)
MCB 301 Spring 2015
Experiment 16: ELISA Lab Notebook
26 points total
Purpose/Relevance (2 points)
Data/Results (4 points)
 Copy of Table 1
Discussion (17 points)
 All results are clearly interpreted, hypotheses are made and evaluated for the obtained
results and their deviation from the “expected” results. There is a good synthesis of the
experiment and the results, particularly in the scope of a larger context. (10 points)
 Which tubes of the mock HIV were the “infected serum” samples? Explain your
reasoning. (2 points)
 Think about the intensity of the reactions with respect to the dilutions on the plate. Is this
information helpful in tracking the infection? Why or why not? (3 points)
 An HIV ELISA should be performed after 6 weeks of initial exposure to HIV. Why? (2
points)
Grammar and Format (3 points)
MCB 301 Spring 2015
Experiment 17: MIC Lab Notebook
24 points total
Purpose/Relevance (2 points)
Data/Results (3 points)
 Completed Tables 1 and 2
Discussion (16 points)
 All results are clearly interpreted, hypotheses are made and evaluated for the obtained
results and their deviation from the “expected” results. There is a good synthesis of the
experiment and the results, particularly in the scope of a larger context. (8 points)
 Using your data, state concisely the effect in E. coli of cat gene dosage (0 copies, one
copy, or multiple copies) on the amount of cellular resistance to chloramphenicol. Your
answer should include a description of the effect on the amount of CAT enzyme
produced and the consequent effect on ribosomal activity. (6 points)
 Does the procedure we used in this exercise determine if an antibiotic is bacteriostatic or
bacteriocidal? Explain your answer. (2 points)
Grammar and Format (3 points)