1 BI 107 Lab Exam 2 Review Questions Exercise 7 Mitosis 1 Define the following term by PoOwbFh

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BI 107, Lab Exam-2 Review Questions
Exercise 7: Mitosis
1. Define the following terms:
Cell cycle, Mitosis, Cytokinesis, Chromosomes, Spindle fibers, Equator plane, Sister-
chromatids, Kinetochore, Centromere, Cell plate and Cleavage furrow.
2. List the different phases of the cell cycle. What happens in each stage?
3. What is the difference between mitosis and cell cycle?
4. List the different phases of mitosis. What happens in each stage? What are the number of
chromosomes and the physical state of chromosomes in each stage?
5. What is cytokinesis? Describe the difference between animal and plant cytokinesis.
6. What is the difference between mitosis and cytokinesis?
Answer all questions in pages 8-9 of Lab Exercise 7
    1. Why are the onion root tip and the whitefish blastula useful tissue for the study of cell
         division?
    2. Distinguish between mitosis and cytokinesis
    3. If a cell has 16 chromosomes when it is in G1, how many chromosomes will there be in
         each daughter cell following a mitotic cell division?
    4. What are the genetic consequences of mitotic cell division for the resulting daughter
         cells?
    5. Why do the lateral and polar views of chromosomes of the same cell in metaphase look
         different?
    6. How do plant and animal cells differ in the execution of cytokinesis? Why don’t plant
         cells undergo cytokinesis in the same manner as animal cells?
    7. List several important values or attributes that mitotic cell division provides to
         multicellular life. In other words, in what ways is mitotic cell division useful for life?
    8. List the 4 stages of the cell cycle and describe the primary cellular activity in each stage?
Very important: You should be able to identify the stages (shown under microscope) of cell
cycle/mitosis in an onion root tip and whitefish blastula. The Math-Science Center (ground floor
of Maclin Tower) has slides and microscopes. If you are not confidant, you may spent more time
on them and study them at your convenience.

Exercise 14: Plasmid Purification and Transformation
Answer all questions in pages 7-10 of Lab Exercise 14
1. Briefly describe the steps of DNA purification.
 Or, What was the rational/logic behind performing each of the following steps in
   plasmid-DNA purification protocol?
   A. Re-suspension of E. coli cells that contain plasmid-DNA.
   B. Addition of cell lysis solution.
   C. Addition of neutralization solution.
   D. Centrifugation after addition of neutralization solution.
   E. Collection of supernatant (the clear fluid).                                                     2
   F. Addition of DNA purification resin to the supernatant
   G. Passing of the mixture of resin and supernatant through the mini-column by
       applying the vacuum.
   H. Washing the column by column wash solution.
   I. Spinning the mini-column after finishing vacuuming.
   J. Addition of 60 microliter water to the mini-column.
   K. Spinning the column after addition of water.

2. What plasmid did you purified in this exercise?
3. What is the link between this plasmid (pAMP) and ampicillin resistance?
4. What is meant by bacterial transformation?
5. How did we know that our transformation experiment worked?
6. What were the controls that we used in our transformation experiment (plates B and C)? Why
did we use these and each control tell us when we compared them to plate A?
7. What is LB Agar?
8. What is pAMP?
9. What is Ampicillin?
10. What is meant by ampicillin resistance?
11. Why the bacteria survive if they accept pAMP from outside? (same as Q.2)

Important: You should be able to interpret the results of a transformation experiment:




         LB Agar                       LB Agar                      LB Agar
        Ampicillin                     Ampicillin                   Plated with
        Plated with                    Plated with                  E. Coli +
        E. Coli +                      E. Coli +                      pAMP
          pAMP                           water



12. Explain what kind of growth you expect to see on each of these plates. Why would we see
this?


Exercise 8 -- Analysis of DNA Using Restriction Enzyme and Electrophoresis:
1. What is nuclease?
2. How does an endonuclease differ from an exonuclease?
3. What is a restriction endonuclease? Write names of some restriction endonuclease.
4. What were 2 restriction endonucleases (RE) that we used in our lab? Write the DNA
sequences these RE recognize. Do they produce sticky ends or blunt ends when they cut the
NDA molecules?
5. How does the number of restriction sites relate to the number of fragments produced for linear
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DNA or circular DNA?
6. What is a palindromic DNA sequence?
7. What is electrophoresis? What does agarose gel electrophoresis allow us to do?
8. What is the chemical nature of agarose?
9. What factors affect the migration rate of DNA through an agarose gel?
10. For DNA molecules of equal sizes, how do the different shapes (conformation) of DNA
differ in terms of distance traveled through an agarose gel?
11. In your DNA electrophoresis experiment, why did you run a DNA ladder (lane 5) and
undigested pAMP DNA (lane 4)?
12. Write some practical applications for use of restriction endonuclease.

13. Examine the following pattern of an electrophoresis gel and Draw a restriction map of
the DNA. Is this DNA circular or linear?


     1        2       3        4       5
                                                         6000 bp
                                                            5000 bp
                                                             4000 bp
                                                            3000 bp
                                                         2000 bp

                                                            1000 bp
                                                               500 bp



                                                         Lane 1: DNA + EcoRI
                                                         Lane 2: DNA + Hind III
                                                         Lane 3: DNA + EcoR I +Hind III
                                                         Lane 4: DNA only
                                                         Lane 5: DNA ladder




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14. Examine the restriction map of plasmid pGEX below.
   A. How many fragments would be produced if the plasmid is digested with Sal I?
      What size would this/these fragment(s) be?
   B. How many fragments would be produced using EcoRI?
     What size would this/these fragment(s) be?
   C. How many fragments would be produced using EcoR I + Hind III + Sal I?
     What size would this/these fragment(s) be?




                                   (4000 end)
                                                  (1 start)
             Sal I (3500 bp)




                                                                     EcoRI
Hind III                       pGEX (4000bp)                         (1000 bp)
(2900 bp)




             Not I (2400 bp)                           EcoRI (1800 bp)


15. Draw the banding pattern for the restriction digestion of the above plasmid as indicated on
right hand side below. The migration of a DNA ladder and its fragment-sizes are identified by
the arrows. .
                                                                                                   5


     1        2        3         4       5
                                                           6000 bp
                                                              5000 bp
                                                               4000 bp
                                                              3000 bp
                                                           2000 bp

                                                               1000 bp
                                                                  500 bp



                                                           Lane 1: DNA + Not I
                                                           Lane 2: DNA + EcoRI
                                                           Lane 3: DNA + EcoR I +Hind III
                                                           Lane 4: DNA+ EcoRI + Hind III + Sal I
                                                           Lane 5: DNA ladder


The following is a real picture of an electrophoresis gel with a particular DNA digested with
EcoR I (Lane 1); HindIII (Lane 2); EcoRI & HindIII (Lane 3); No enzyme (Lane 4). Lane five
contains DNAs of known sizes.

 1       2        3    4     5




                                                5000 bp
                                                4000 bp
                                                3000 bp

                                                2000 bp
                                                1500 bp
                                                1000 bp

                                                500 bp



Estimate the size of the DNA molecule. Is this DNA molecule circular or linear? Draw a
restriction map of the DNA at the right hand side of the picture.
16. Examine the following gel. Determine whether the DNA is prokaryotic or eukaryotic. Draw a
restriction map of the DNA.

                                                8000 bp
       1      2      3      4     5

                                               7000 bp
                                               7000 bp

                                                6000 bp

                                               5000bp

                                                4000 bp

                                                3000 bp

                                                 2000 bp
                                                 1000 bp

                                                  500 bp


                                                        Lane 1: DNA + EcoRI
                                                        Lane 2: DNA + Hind III
                                                        Lane 3: DNA + EcoR I +Hind III
                                                        Lane 4: DNA only
                                                        Lane 5: DNA ladder
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Exercise 11: Plant Cells, Tissues and Organs
1. Define tissue
2. Define Organ
3. Identify root system, shoot system, stem, leaves, node, internode, blade and periole.
4. How does a tap root differ from a diffuse root?
5. What are the four major plant tissues? Describe their functions.
6. Distinguish between a longitudinal section and a cross section.
7. Identify the different parts of a longitudinal section of a growing root.
   A. What is the function of root cap? What kind of tissue is the root cap made of?
   B. What is the function of region of cell division? What kind of tissue is this region
      made of?
   C. What is the function of the root hairs?
   D. What is the function of apical meristem?
8. Identify and describe the functions of different parts of cross section of a root:
   epidermis, cortex, vascular cylinder, xylem, phloem, endodermis, parenchyma cells.
9. Identify and describe the functions of different parts of cross section of a woody dicot
  stem: pith, xylem, annual ring, vascular ray, vascular cambium, phloem, cortex
   periderm.
10. What can we tell about from the annual ring?
11. How are the annual rings formed?
12. Identify and describe the functions of different parts of a leaf cross section: upper
     epidermis, lower epidermis, guard cell, stomata, veins, palisade layer, spongy layer,
     mesophyll.
13. In which epidermal layer are stomata more prevalent?
14. What types of gases move in and out of the leaf through the stomata?
Also answer the questions in your lab manual.

Exercise 12: Animal Cells and Tissues
1. Describe the functions of the 4 types of animal tissue.
2. Identify and describe the functions of different tissues and structures in frog skin:
 a) poison glands b) epidermis c) dermis e) mucus gland f) lumen of poison gland and
 mucus gland g) chromatophores h) stratified squamous epithelium of the epidermis
 i) cuboidal epithelium of mucus and poison gland.
3. Identify and describe the functions of following tissues and structures of trachea:
   pseudostratified ciliated columnar epithelium, goblet cells, cartilage, chondrocytes,
   matrix.
4. What is the matrix of cartilage made of?
5. Identify and describe the functions of Haversial system, Haversian canal, osteocytes,
canaliculi, lacuna, matrix in bone.
6. What are the major cells in blood?
7. Identify and describe the functions of erythrocytes, leucocytes, neutrophils, lymphocytes in
blood.
8. Identify muscle fibers, striations, nuclei in a cross section of striated muscle.
9. What is the function of striated muscle?
10. Identify and describe the functions of different cells in nervous tissue.
11. Identify the following parts of a neuron: cell body, axon, dendrites, nucleus.
Restriction Mapping: An example

								
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