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I have left up last semester's review sheet as some students find these useful.
It will be updated for Fall 2010 2-5 days before the exam.
BIOL360 Lab Exam 2 Review sheet Fall
2009
My little disclaimer:
This exam review sheet is intended to help you
prepare for the exam but should not be considered your only source of study. It is
possible I have inadvertently left off some items of importance, so you should use your
notes and lab handouts to help you prepare for the exam. You are still responsible for any
material we covered in lab, whether on this sheet or not.
The lab will consist of 50-60 multiple choice questions.
There will be a written exam much like last time, and questions more like a
traditional lab practical with certain materials setup for you to answer questions
about. The exam will start
promptly at 2:00 and will end at
3:30. Students arriving late will not be given additional
time. Please bring #2 pencils.
Human Chromosome Analysis/Karyotyping Lab
 | Be able to match up homologous pairs of chromosomes (from a limited
sample). |
 | Be able to distinguish between chromosomes on the slides (or photograph), given
information about them. (As you did in the online
lab, using the chromosome
descriptions) |
 | Be able to diagram and label (centromere, p arm, q arm) an indicated chromosome
under the microscope (or photograph).
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 | Review telocentric, metacentric, etc.
|
 | Be familiar with the karyotyping procedure
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 | Be able to recognize the karyotype of common
disorders. |
 | Review Pedigrees and the symbols used |
 | What would a pedigree of a dominant disorder look
like? A recessive disorder? A sex-linked disorder? |
Use of micropipette/ Gel electrophoresis
 | Be able to answer some questions about routine micropipettor use |
 | Be able to tell what volume is indicated on a particular micropipette |
 | How is a gel formed? What is it made out of? |
 | What is the purpose of the
electrophoresis buffer? |
 | How is DNA separated by gel electrophoresis? (Consider the charge of DNA and the
size of DNA) |
 | How is a gel actually run? |
 | What is a comb? casting tray?
electrophoresis chamber? Power supply? |
 | What is a typical voltage for running
the gels? |
 | What was done to the gel to visualize the DNA
in the gel? (For DNA isolation lab
and for Cancer gene detection lab) |
 | Be able to name indicated parts of an electrophoresis setup.
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DNA isolation Lab
 | Review the basic steps involved in isolation of DNA from cells. What is the
purpose of sarkosyl, protease, and EDTA in this extraction procedure? |
 | Review questions at end of exercise. |
 | Be able to distinguish between degraded and nondegraded DNA on a gel. |
 | What type of DNA was isolated? (plasmid or chromosomal?) |
 | What is the purpose of the glycerol in the loading
dye? What is the purpose of the blue dye in the loading dye? |
 | What special precautions have to be taken when isolating this type of DNA?
Why? |
 | What organism was the DNA isolated from? |
 | How did we stain the gel to visualize the DNA? (read
the lab handout--this was done after some of you left) |
Bacterial Transformation Lab
 | What plasmid was used? |
 | What special features does it have? |
 | What genes are on the plasmid? what do they do? |
 | Be familiar with the entire transformation procedure
 | how are cells made competent? (what is
meant by this term?) |
 | what steps are taken in the procedure
and in what order? |
 | What temperatures are involved? |
 | what sort of plates were the
transformed cells plated on? |
|
 | |
 | Know the characteristic of the 4 plates you made: -DNA/LB, -DNA/LB/amp,
+DNA/LB/amp, and +DNA/LB/amp/ What should you see
(in terms of bacterial growth) on each plate |
 | Which plates should be compared to determine if any genetic transformation has
occurred? Why? |
 | What is the role of IPTG in the plates? |
 | What is transformation efficiency? How do you calculate transformation
efficiency? Was ours good? |
 | Be able to answer questions about actual transformation plates.
Be able to calculate transformation efficiency. |
 | How would you select for cells with plasmid? How could you distinguish
between cells with non-recombinant plasmid and recombinant plasmid? |
 | Review questions at end of exercise. |
Cancer Cell Morphology/Cancer Gene Detection Lab
 | review distinguishing features of cancer cells:
 | immortal |
 | loss of contact inhibition (cells overgrow and pile up in culture) |
 | loss of anchorage independence (cells in culture are less adherent to one another
and to tissue culture plate) |
 | altered cell morphology |
|
 | Review cell staining procedure . What
stains are used? |
 | Which stain colored the cytoplasm? which stained the
DNA? |
 | Be able to distinguish between normal and cancerous cells on a prepared slide |
 | germline vs. somatic mutation |
 | Be able to distinguish between a hereditary and a sporadic cancer based on a
family pedigree |
 | What is Li-Fraumeni syndrome? What mutation do they carry? |
 | review information on p53 from handout |
 | review procedure-- what did we do? Why? |
 | why did we heat the DNA right before loading on the
gel? See p. 14 of handout |
 | What was done to the DNA prior to loading it on the gel (by the company that sent
the DNA). See p. 10 of your handout. |
 | Be able to answer questions about the gel, and what the different bands represent |
 | How were Valerie's children tested to determine if
they had a gene mutation? |
 | Review questions at end of exercise. |
 | Normal cells:
 | have defined lifespan in culture. They will grow for a while, undergo
"crisis" and then die |
 | contact inhibited -- cell growth ceases under conditions of decreased nutrients,
injury, or cell division, or if space is physically limiting (crowding on plate) |
 | cells adhere to tissue culture plate and to one another
|
|
 | Cancer cells have distinguishing characteristics:
 | immortal |
 | loss of contact inhibition - cells overgrow, pile on top of one another |
 | anchorage independent -- cancer cells are less adherent to each other and the
tissue culture plate |
 | morphological changes |
 | chromosomal aberrations |
|
 | All types of cancer have a common distinguishing feature:
LOSS OF GROWTH CONTROL |
 | Proto-oncogenes
 | normal genes, encode normal proteins involved in accelerating growth of the cell |
 | if mutated, become oncogenes |
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 | Tumor suppressors
 | normal function--restrict cellular proliferation |
 | if mutated or inactivated, cellular proliferation without regulation |
 | Review information on p53 -- what type of protein is it? What is its normal
function? |
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