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Documente Cultură
by Dr. Rego
Objective: In this lab you will learn about and examine the underlying processes required for cell
division (the cell cycle).
Instructions for Students: Read the entire laboratory procedure and complete all prelab activities
before coming to lab. Once you come to lab follow the steps provided for each exercise, take good
notes and make sure to get checked off by the lab instructor once you finish. Your instructor will let
you know if you are required to complete any additional activities in for lab.
Learning Objectives: After completing this laboratory experiment, you should be able to do the
following things:
1.
2.
3.
4.
Figure 1. The various steps in the mitotic cell cycle. The mitotic cell cycle begins once a somatic
cell receives cues to divide and results in the production of 2 identical daughter cells.
o Interphase
Interphase can be thought of as a preparatory phase for mitosis. In order for a cell to produce 2
identical copies of itself, the material inside the cell must double. This includes cytoplasmic
material, such as organelles (e.g. the mitochondria), as well as nuclear material like DNA. In the 1st
stage of interphase (G1) the cell grows and duplicates much of its cytoplasmic material. The next
stage of interphase (S), known as the synthesis phase, is where cells replicate their DNA leading to
the production of enough genetic material for 2 cells. Additional growth occurs in the last stage of
interphase (G2). It is important to note that the genetic material of cells remains in the nucleus and
has not yet condensed during interphase, therefore, we would not expect to see any visible
chromosomes during this stage of the cell cycle. In fact, it is important that the DNA is spread out
during interphase because it is much easier to replicate DNA that is spread throughout the nucleus
compared to DNA that is condensed into chromosomes.
o Mitosis
Mitosis means the division of the nucleus to produce two identical daughter cells. As cells pass from
interphase to prophase (Figure 2A) the chromosomes condense and the mitotic spindle forms. At
this point each chromosome is composed of two sister chromatids joined at the centromere.
Additionally, the nuclear envelope breaks down and the mitotic spindle begin to attach to the
chromosomes at the centromere. During metaphase (Figure 2B) the chromosomes line up in the
middle of the cell forming a structure called the metaphase plate. This stage is important to ensure
that DNA is split equally so each daughter cell receives the appropriate amount of genetic material.
During anaphase (Figure 2C) the centromere splits and each chromatid, now a chromosome, is
pulled to opposite sides of the cell. In the last stage, telophase (Figure 2D), the chromosomes
become less condensed, two new nuclei form and the mitotic spindle disintegrate. This officially
ends mitosis which, as mentioned before is the division of the nucleus. The cell cycle ends with
cytokinesis, the division of cytoplasm into 2 daughter cells, which often overlaps with late
telophase.
Now you have 2 pairs of homologous chromosomes. Remember that in each cell of
your body, you have 2 copies of each chromosome (1 from mom and 1 from dad).
The red can represent mom and the yellow can represent dad. The 2 long
chromosomes (homologous chromosomes) will code for the same traits, for
example eye color, hair color etc. The 2 short chromosomes will also code for the
same traits. However, these traits are different from the traits coded for by the long
chromosomes. The difference between the red long chromosome and the yellow
long chromosome is that they may contain different alleles. For example, if the eye
color trait is found on the long chromosome you may have received a copy of the
blue eye allele from your mom (red bead 3) and a copy of the brown allele at the
same position on the other chromosome from your dad (yellow bead 3).
2. Replicate your chromosomes to make 4 replicated chromosomes
a. Perform the same procedure as step one to make 2 more long and 2 more short
chromosomes
b. Connect the identical chromosomes together by touching their magnet centromeres
together
At the point you may think you have 8 chromosomes, however, you still only have 4
chromosomes but in this case they are replicated. During replication, you copied the
original 4 chromosomes but did not make any new chromosomes. In other words,
before replication, you had 4 chromosomes but after replication you still have 4
chromosomes, each consisting of 2 chromatids joined together at the centromere.
Now that we have replicated our chromosomes and have twice as much DNA as we
need for a normal cell, we can begin to simulate the process of mitosis!
Critical thinking question: What is the difference between homologous chromosomes (Step 1)
and sister chromatids (Step 2)?
3. Prophase
a. We dont need to do anything with the beads but in the blanks below list the 3 things
that happen to a cell during prophase
i. _____________________________________________________________________________
ii. _____________________________________________________________________________
iii. _____________________________________________________________________________
4. Metaphase
a. Line up your chromosomes as they would be found in metaphase (in a straight
vertical line)
b. Attach the spindles to each chromatid
i. Cut 8 pieces of string and tie them to the centromere of each chromatid
c. Place the ends of the strings near the centriole away from the middle of your cell
5. Anaphase
a. Pull the chromosomes apart away from the middle and towards the centrioles
6. Telophase
a. Recognize how many chromosomes you end up with in EACH daughter cell
b. Draw each step of mitosis you recapitulated with your model in the diagram below:
Procedure:
1. Obtain an onion root tip slide from the slide holder
2. Place your slide on the microscope stage
3. Observe the onion root tip under low power
a. Cells at the very tip of an onion are not dividing, therefore, focus on the cells just
behind the tip (this is known as the zone of cell division)
4. Once you have located the zone of cell division increase the power of magnification until
you can clearly see individual cells
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5. Try to locate cells from each stage of mitosis and draw below:
meiosis I occurs in Prophase I. During prophase I the homologous chromosomes pair up and
exchange genetic material, a process known as crossing over. The process of exchange actually
occurs between the chromatids of the homologous chromosomes (Figure 3). This exchange of
genetic material increases the genetic variation in the offspring. The next stage, metaphase I, is
where chromosome pairs line up next to each other in the middle of the cell. The way the
chromosomes line up is random meaning that there is a 50:50 chance that a chromosome lines up
on one side or the other (Figure 4 Law of independent assortment). The next difference between
mitosis and meiosis occurs in
anaphase I. During anaphase I the
homologous chromosomes
segregate and move to the
opposite poles. The cells then
enter a second cell cycle (meiosis
II) without replicating DNA. The
length of interphase between
meiosis I and meiosis II varies from nonexistent too years depending on the organism. In meiosis II
during Anaphase II the centromere divides and the sister chromatids are pulled to opposite poles of
the cell.
Procedure:
1. Build chromosomes for a diploid cell (2n) with 4 un-replicated chromosomes
a. Obtain 4 magnetic centromeres
b. Make 2 homologous long chromosomes
i. Connect 2 sets of 3 yellow beads to 1 centromere
ii. Connect 2 sets of 3 red beads to another centromere
c. Make 2 homologous short chromosomes
i. Connect 2 sets of 1 yellow bead to 1 centromere
2.
3.
4.
5.
6.
7.
8.
9.
10.
Procedure:
1. Obtain a grasshopper testis slide from the slide holder
2. Place your slide on the microscope stage
3. Observe the grasshopper testis under low power
4. Once you have focused the sample under low power increase the power of magnification
until you can clearly see individual cells
5. Try to locate cells from each stage of meiosis and draw below:
a. For those stages you cant find please draw what you would expect to see
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Summary questions:
1. List the steps in mitosis and meiosis in the table below and identify how these processes
differ.
Mitosis
Meiosis
2. Describe crossing over and explain how crossing over contributes to genetic variability?
How is this beneficial?
3. How does the cell decide which homologue goes to which pole during anaphase I of
meiosis?
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