AP® Investigation #7
Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Table of Contents
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Abstract. . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
General Overview . . . . . . . . . . . . . . . . . . . . . . 1
Recording Data. . . . . . . . . . . . . . . . . . . . . . . . 2
Material Requirements/Checklist . . . . . . . . . . . . . . 4
curriculum alignment . . . . . . . . . . . . . . . . . . . . 5
Learning Objectives. . . . . . . . . . . . . . . . . . . . . . 6
Time Requirements . . . . . . . . . . . . . . . . . . . . . . 5
Safety Precautions. . . . . . . . . . . . . . . . . . . . . . 7
Pre-Lab Preparations. . . . . . . . . . . . . . . . . . . . . 8
Student guide contents
Background. . . . . . . . . . . . . . . . . . . . . . . 11
Part 1: Cell Size & Diffusion. . . . . . . . . . . . . . . 13
Part 2: Modeling Osmosis in Living Cells. . . . . . . . 17
Part 3: Osmosis in Living Plant Cells . . . . . . . . . . 21
Assessment Questions/Additional Questions (Optional)24
MATERIAL SAFETY DATA SHEETS. . . . . . . . . . . . . . . . . .
**AP® and the Advanced Placement Program are registered trademarks
of the College Entrance Examination Board. The activity and materials
in this kit were developed and prepared by WARD’S Natural Science
Establishment, which bears sole responsibility for their contents..
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
abstract
In this lab, students will examine and compare the phases of mitosis and meiosis in plant and animal
cells. Students will then determine the relative time cells spend in each phase and calculate the
distance between a specific gene and the chromosome centromere. Students will accomplish this by
preparing slides, making observations using a compound microscope, and manipulating data.
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general Overview
The College Board has revised the AP Biology curriculum to begin implementation in the fall of
2012. Advanced Placement (AP) is a registered trademark of the College Entrance Examination
Board. The revisions were designed to reduce the range of topics covered, to allow more depth of
study and increased conceptual understanding for students. There is a shift in laboratory emphasis
from instructor-designed demonstrations to student-designed investigations. While students may be
introduced to concepts and methods as before, it is expected that they will develop more independent
inquiry skills. Lab investigations now incorporate more student-questioning and experimental
design. To accomplish this, the College Board has decreased the minimum number of required
labs from 12 to 8 while keeping the same time requirement (25% of instruction time devoted to
laboratory study). The College Board has defined seven science practices that students must learn to
apply over the course of laboratory study. In brief, students must:
1. Use models
2. Use mathematics (quantitative skills)
3. Formulate questions
4. Plan and execute data collection strategies
5. Analyze and evaluate data
6. Explain results
7. Generalize data across domains
The College Board published 13 recommended laboratories in the spring of 2011. They can be found
at: http://advancesinap.collegeboard.org/science/biology/lab
Many of these laboratories are extensions of those described in the 12 classic labs that the College
Board has used in the past. The materials provided in this lab activity have been prepared by
Ward’s to adapt to the specifications outlined in AP Biology Investigative Labs: An Inquiry-Based
Approach (2012, The College Board). Ward’s has provided instructions and materials in the AP
Biology Investigation series that complement the descriptions in this College Board publication.
We recommend that all teachers review the College Board material as well as the instructions here
to get the best understanding of what the learning goals are. Ward’s has structured each new AP
investigation to have at least three parts: Structured, Guided, and Open Inquiry. Depending on a
teacher’s syllabus, s/he may choose to do all or only parts of the investigations in scheduled lab
periods.
The College Board requires that a syllabus describe how students communicate their experimental
designs and results. It is up to the teacher to define how this requirement will be met. Having
students keep a laboratory notebook is one straightforward way to do this.
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Page Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Recording Data in a Laboratory Notebook
All of the Ward’s Investigations assume that students will keep a laboratory notebook for studentdirected investigations. A brief outline of recommended practices to set up a notebook, and one
possible format, are provided below.
1. A composition book with bound pages is highly recommended. These can be found in most
stationary stores. Ward’s offers several options with pre-numbered pages (for instance, item
numbers 32-8040 and 15-8332). This prevents pages from being lost or mixed up over the
course of an experiment.
2. The title page should contain, at the minimum, the student’s name. Pages should be numbered in
succession.
3. After the title page, two to six pages should be reserved for a table of contents to be updated as
experiments are done so they are easily found.
4. All entries should be made in permanent ink. Mistakes should be crossed out with a single line
and should be initialed and dated. This clearly documents the actual sequence of events and
methods of calculation. When in doubt, over-explain. In research labs, clear documentation may
be required to audit and repeat results or obtain a patent.
5. It is good practice to permanently adhere a laboratory safety contract to the front cover of the
notebook as a constant reminder to be safe.
6. It is professional lab practice to sign and date the bottom of every page. The instructor or lab
partner can also sign and date as a witness to the veracity of the recording.
7. Any photos, data print-outs, or other type of documentation should be firmly adhered in the
notebook. It is professional practice to draw a line from the notebook page over the inserted
material to indicate that there has been no tampering with the records.
For student-directed investigations, it is expected that the student will provide an experimental plan
for the teacher to approve before beginning any experiment. The general plan format follows that of
writing a grant to fund a research project.
1. Define the question or testable hypothesis.
2. Describe the background information. Include previous experiments.
3. Describe the experimental design with controls, variables, and observations.
4. Describe the possible results and how they would be interpreted.
5. List the materials and methods to be used.
6. Note potential safety issues.
(continued on next page)
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Recording Data in a Laboratory Notebook (continued)
After the plan is approved:
7. The step-by-step procedure should be documented in the lab notebook. This includes recording
the calculations of concentrations, etc., as well as the weights and volumes used.
8. The results should be recorded (including drawings, photos, data print outs, etc.).
9. The analysis of results should be recorded.
10. Draw conclusions based on how the results compared to the predictions.
11. Limitations of the conclusions should be discussed, including thoughts about improving the
experimental design, statistical significance, and uncontrolled variables.
12. Further study direction should be considered.
The College Board encourages peer review of student investigations through both formal and
informal presentation with feedback and discussion. Assessment questions similar to those on the AP
exam might resemble the following questions, which also might arise in peer review:
•
Explain the purpose of a procedural step.
•
Identify the independent variables and the dependent variables in an experiment.
•
What results would you expect to see in the control group? The experimental group?
•
How does XXXX concept account for YYYY findings?
•
Describe a method to determine XXXX.
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Page Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Materials checklist
MATERIALS PROVIDED IN KIT
MATERIALS NEEDED BUT NOT PROVIDED
Units
per kit
Description
Garlic clove
8
Scalpels, disposable
Compound microscope
1 box 72
Precleaned microscope slides,
Absorbent wipes
1 box 100
Razor blades
Glass marking pens
8
22 mm plastic coverslips,
Live materials coupon for
Sordaria cross demo plate*
Large wooden clothespin,
4
Prepared slide, fish mitosis
Ruler
4
Prepared slide, onion root mitosis
Toothpicks
15
Pipets, 6’’ grad.
Box (or dark area to grow roots)
1
Hydrochloric acid, 1 M, 30 mL,
Dissection scissors
1
Carbol fuchsin solution, 30 mL
Timer
8
Disposable inoculating loop
Forceps
600
Red pop beads
Bunsen burner
600
Pink pop beads
Goggles, aprons, and gloves
1
Sand
32
Magnetic yellow centromeres
32
Centrioles, clear
OPTIONAL MATERIALS (NOT PROVIDED)
Prepared sordaria squash slides (912200) or squash
cards (323377)
NaC
1
70% isopropyl alcohol, 30 mL,
pH 3 buffer and/or pH 10 buffer 1
Instructions (this document)
Lectin
(phytohemmaglutinin PHA-m, increases mitosis)
1
100 mL beaker
Uric acid (decreases mitosis)
NaCl
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* - It is recommended that you
redeem your coupon for live/
perishable materials as soon as
possible and specify your preferred
delivery date. Generally, for timely
delivery, at least a week’s advance
notice is preferred.
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Page Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
This lab activity is aligned with the 2012 AP Biology Curriculum (registered trademark of the College Board).
Listed below are the aligned Content Areas (Big Ideas and Enduring Understandings), the Science Practices, and the
Learning Objectives of the lab as described in AP Biology Investigative Labs: An Inquiry Approach (2012). This is a
publication of the College Board that can be found at http://advancesinap.collegeboard.org/science/biology/lab.
Curriculum alignment
Big Idea
‹ Big Idea 3: Living systems store, retrieve, transmit, and respond to information essential to life
processes.
Enduring Understandings
‹ 3A1: DNA, and in some cases RNA, is the primary source of heritable information.
‹ 3A2: In eukaryotes, heritable information is passed to the next generation via processes that
include the cell cycle and mitosis or meiosis plus fertilization.
‹ 3A3: The chromosomal basis of inheritance provides an understanding of the pattern of passage
(transmission) of genes from parent to offspring.
‹ 3C2: Biological systems have multiple processes that increase genetic variation.
Science Practices:
‹ 1.2 The student can describe representations and models of natural or man-made phenomena and
systems in the domain.
‹ 5.3 The student can evaluate the evidence provided by data sets in relation to a particular scientific
question.
‹ 6.2 The student can construct explanations of phenomena based on evidence produced through
scientific practices.
‹ 6.4 The student can make claims and predictions about natural phenomena based on scientific
theories and models.
‹ 7.1 The student can connect phenomena and models across spatial and temporal scales.
‹ 7.2 The student can connect concepts in and across domain(s) to generalize or extrapolate in and/
or across enduring understandings and/or big ideas.
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Page Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
learning objectives
‹ The student can make predictions about natural phenomena occurring during the cell cycle (3A2
& SP 6.4).
‹ The student can describe the events that occur in the cell cycle (3A2 & SP 1.2).
‹ The student is able to construct an explanation, using visual representations or narratives, as to
how DNA in chromosomes is transmitted to the next generation via mitosis, or meiosis followed
by fertilization (3A2 & SP 6.2).
‹ The student is able to represent the connection between meiosis and increased genetic diversity
necessary for evolution (3A2 & SP 7.1).
‹ The student is able to evaluate evidence provided by data sets to support the claim that heritable
information is passed from one generation to another generation through mitosis, or meiosis
followed by fertilization (3A2 & SP 5.3).
‹ The student is able to construct a representation that connects the process of meiosis to the
passage of traits from parent to offspring (3A3 & SP 1.1, SP 7.2).
‹ The student is able to construct an explanation of the multiple processes that increase variation
within a population (3C2 & SP 6.2).
Time Requirements
Part 1: Mitosis (Structured Inquiry)
1a: Modeling Mitosis
20 minutes
1b: Onion and fish prepared slides
45 minutes
1c: Garlic Root Tip Squash
45 minutes
Part 2: Meiosis (Guided Inquiry)
20 minutes*
2a: Modeling Meiosis
* – Optional: This part may be done alongside Part 1a to save time.
2b: Meiosis & Sordaria Crossing Over
45 minutes
Part 3: Open Inquiry
NOTE: If you choose to have chemicals that
modulate mitosis available for student directed
inquiry, order these far enough in advance to
assure they arrive prior to the start of the lab.
Students will have to re-grow garlic roots as part
of their experiments.
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Varies, depending on students’ experimental designs.
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Page Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Safety Precautions
Lab-Specific Safety
‹ 70% isopropyl alcohol is poisonous if ingested, and will irritate the eyes. Wear safety goggles.
Read the MSDS for this chemical.
‹ Hydrochloric acid is a mild irritant. Avoid contact with the skin and eyes. Poisonous if ingested.
Read the MSDS for this chemical.
General Safety
‹ The teacher should be familiar with safety practices and regulations in their school (district and
state). The teacher should know what needs to be treated as hazardous waste and how to properly
dispose of non-hazardous chemicals or biological material.
‹ Consider establishing a safety contract that students and their parents must read and sign off on.
This is a good way to identify students with allergies to things like latex so that you (and they)
will be reminded of what particular things may be risks to individuals. A good practice is to
include a copy of this contract in the student lab book (glued to the inside cover).
‹ Students should know where all emergency equipment (safety shower, eyewash station, fire
extinguisher, fire blanket, first aid kit etc.) is located.
‹ Make sure students remove all dangling jewelry and tie back long hair before they begin.
‹ Remind students to read all instructions, Material Safety Data Sheets (MSDSs) and live care
sheets before starting the lab activities and to ask questions about safety and safe laboratory
procedures. Appropriate MSDSs and live care sheets can be found on the last pages of this
booklet. Additionally, the most updated versions of these resources can be found at
www.wardsci.com, under Living Materials http://wardsci.com/article.asp?ai=1346.
(Note that in this particular lab, there are no live materials that require a live care sheet.
‹ In student directed investigations, make sure that collecting safety information (like MSDSs) is
part of the experimental proposal.
‹ As general laboratory practice, it is recommended that students wear proper protective
equipment, such as gloves, safety goggles, and a lab apron.
At end of lab:
‹ All laboratory bench tops should be wiped down with a 20% bleach solution or disinfectant to
ensure cleanliness.
‹ Remind students to wash their hands thoroughly with soap and water before leaving the
laboratory.
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Page Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Pre-Laboratory Preparation
Notes
1 week before lab:
Redeem your coupon for your Sordaria demonstration cross plate
at least one week prior to starting the lab to allow sufficient time for
delivery. If the line of growth between the agar cubes appears dark
and well-developed, store the plate in a refrigerator, agar side up, until
ready to use. If the line of growth appears to be light, incubate at room
temperature until it resembles the plate in Figure __________.
1 week - 2 days before lab:
Prepare the Garlic Root Tips for Part 1c: Garlic Root Tip Squash
‹
If time is a concern, you may grow the garlic root tips in
advance. You may also have your students grow the root
tips; the following steps are also included in the students’
procedure.
1. Obtain a jar or Petri dish.
2. Using a ruler, measure 1.5 cm starting at the bottom of the
container and mark the measurement with a permanent marker.
3. Fill the jar with the fine sand to the mark on the jar. You may
need to sift or swirl the sand back and forth to ensure you have
distributed the sand evenly.
4. Add water to the jar to wet the sand.
5. Separate a clove of garlic into individual sections, or “toes”.
Remove the paper-like skin from each section and any dried
roots on the primordia (blunt end) with a razor blade.
‹
The section used must have root primordia present or it
will not produce root tips. The root tips will grow within
2 days for very fresh garlic, or three days for older garlic.
Because garlic has a mitotic cycle of approximately 12.5
hours, it’s important to “plant” the garlic and harvest
the root tips at approximately the same time of day in
order to get the greatest percentage of meristematic cells
undergoing mitosis.
6. Place the cloves into the container with the root primordia (blunt
end) in the sand.
(continued on next page)
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Page Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Pre-Laboratory Preparation (continued)
Notes
7. Place the jar in a box or dark place until the root tips have grown
to a length of about 4 to 5 mm. Several viable root tips will
grow on each section of garlic.
‹
It is important that the garlic grows in the dark to ensure that
it produces roots rather than shoots.
8. Remove the garlic from the box approximately one half-hour
before performing the experiment to expose the root tips to
light.
‹
2 to 3 mm of the root tip can be cut off and stored in 70%
isopropyl alcohol for up to 1 week.
Immediately before lab:
1. Make copies of Student Guide.
Copy pages __ to __ of the student copymaster prior to starting
class.
2. Count out beads.
In preparation for modeling mitosis (Part 1a) or meiosis (Part
2a), you may count out the beads for each group. Provide each
lab group with the following:
•
40 red pop beads
•
40 pink pop beads
•
4 yellow magnetic centromeres
•
4 clear plastic centrioles
Extension Activity: Loss of cell cycle control
Obtain photos of normal and Philadelphia type leukemia
chromosomes. If time permits, students may obtain these via computer
searches. This can also be done as part of a classroom or homework
assignment.
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Page 10
Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Background
OBJEcTIVES
‹ Make predictions about natural
phenomena occurring during the
cell cycle,
‹ Describe the events that occur
in the cell cycle.
‹ Construct an explanation,
using visual representations or
narratives, as to how DNA in
chromosomes is transmitted
to the next generation via
mitosis, or meiosis followed by
fertilization.
‹ Represent the connection
between meiosis and increased
genetic diversity necessary for
evolution.
‹ Evaluate evidence provided by
data sets to support the claim
that heritable information is
passed from one generation
to another generation through
mitosis, or meiosis followed by
fertilization.
‹ Construct a representation that
connects the process of meiosis
to the passage of traits from
parent to offspring.
‹ Construct an explanation of the
multiple processes that increase
variation within a population..
Reproduction in living things can be achieved through either asexual
or sexual processes. In asexual reproduction, offspring organisms
are genetically similar to the parent organism with limited, if any,
introduction of DNA from another individual. In sexual reproduction,
offspring organisms are generated by combining one half of the
DNA from each of two parent individuals to produce a diploid (2N)
offspring that has two complete sets of chromosomes and genes. The
process of sexual reproduction enables greater genetic variation in
offspring than asexual reproduction.
Multicellular, diploid, eukaryotic organisms use both types of cellular
reproduction. Growth and development of the organism occurs through
asexual reproduction of cells through the process of mitosis where
every normal daughter cell is diploid, carrying two copies of each
chromosome and set of genes. In mitotic reproduction, these daughter
cells are genetic replicates of both the parent cell and the sister cell.
Most single cell eukaryotes reproduce by simple mitosis, producing
more replicates of the parent organism.
Mitosis
The mitotic cell cycle is a highly regulated process that insures
accuracy in DNA replication and equal division into daughter
cells. Defects in accurate reproduction will cause abnormalities that
will result in decreased viability of both the daughter cells and the
multicellular organism through a variety of mechanisms (including
apoptosis and cancer).
Mitosis is a continuous process that can be described by a sequence
of stages. The stages are defined by molecular and cytosolic events.
Visual inspection of dividing cells through the microscope has defined
three main stages (Figure 1) that can be further subdivided (Figures 2,
3 on the next pages):
‹ please check figure #’s - these were
renumbered, for better clarity
Mitosis:
The cells are separating DNA equally for two future daughter
cells.
Cytokinesis:
The daughter cells are separated into two individual cells,
Figure 1
Interphase:
The cells look quiescent,
(continued on next page)
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Background (COntinued)
Notes
In interphase, the cell may be arrested in this part of the cell cycle
(referred to as the G0 part of interphase) as a diploid cell (2N). If the
cell continues through the next mitotic cycle, interphase only appears
to be quiescent. This is when the cell grows in preparation of another
cycle (G1 ), DNA is replicated (S phase, cell becomes 4N), the DNA
checked for errors and repaired prior to mitosis (G2).
Figure 2: Interphase cell in onion root tip.
Interphase
Prophase Metaphase
Anaphase
Telophase Cytokinesis
Figure 3:
Visually distinct stages of the mitotic cycle in the onion root tip.
Mitosi s is subdivided into four stages that can be identified through
the microscope:
Figure 4: Prophase –early, in onion root tip,
•
The replicated DNA condenses around histone proteins and
supercoils to form visible chromosomes
•
The nuclear membrane dissolves
•
Centioles form and begin to migrate to opposite sides of the cell
•
Microtubules organize around the centrioles to form mitotic
spindles
•
Spindle fibers attach to the kinetochore proteins at the centromere
(holding sister chromotids together).
(continued on next page)
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Background (COntinued)
Notes
Figure 5: Metaphase
• Centrioles complete migration to opposite poles of the
cell
•
Sister chromatids line up along the midline of the cell
Figure 6: Anaphase
•
Spindles shorten, pulling sister chromotids apart to opposite poles
of the cell
Figure 7: Telophase
•
Chromosomes de-condense
•
2 nuclear envelopes form around the separated chromosomes
(each nucleus is 2N again)
Cytokensis generally follows and divides the cytosol and cellular
membrane to yield two cells that are identical. In plant cells, small
vesicles move along microtubules to the mid line. These vesicles fuse
to form a cell plate, which grows to form the cell wall that separates
the cells.
Control of the cell cycle
To maintain normal growth and development it is essential that cells
divide only when and where needed. Therefore, the transition between
the stages of the cell cycle is a tightly controlled process. Levels
of of proteins called cyclins build up in the cell and bind to cyclindependent kinase, forming CDK complexes. These CDK molecules
either add or remove phosphate groups from substrates to move the
cell to the next stage of the cycle. In order to progress, however, the
cell must pass through several “checkpoints.” These checkpoints
assure that cells only divide when needed and when DNA duplication
is completed and without errors.
(continued on next page)
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Background (COntinued)
Notes
The three main checkpoints are:
Restriction Point – this occurs in G1. After division the daughter cells
will continue to grow in size for a short time but are halted at the
restriction point. Here cells will either terminally dif ferentiate and
enter what is termed G0, or the cells will be stimulated a by growth
factor and continue through the cycle again.
G2/M Checkpoint - occurs between the end of G2 and the beginning
of mitosis. Specialized molecules read the newly formed DNA and
will delay the cell from entering mitosis if there are strand breaks or
if inappropriate nucleotides are incorporated. The cell cycle will only
continue if repairs can be made, otherwise, the cell will die without
completing the cycle.
Metaphase/Anaphase Checkpoint - early in mitosis. Specialized
proteins on each centromere, called the kinetochores, activate if
microtubules are attached and appropriate force is being applied.
Activation allows the chromosomes to separate. If activation does not
occur, mitosis will halt and the cell will die without completing the cell
cycle.
Mutations in cells that lead to the loss of cell cycle control result in a
loss of control over growth, cell differentiation, and death. Mutations
may accumulate over time causing the cell clones to become a tumor;
the tumor may later become metastatic if the cell acquires the ability to
establish separate masses in distant tissues. These cancerous mutations
usually affect genes which encode proteins involved in the control of
other genes (transcriptional regulation), cellular metabolism (speeding
up or slowing down cell growth), and/or the cell cycle of division and
growth. Also affected are hormones and their receptors, regulatory
molecules such as cytokines and their receptors, and DNA repair
mechanisms.
‹ referecne to figure 8 somewhere
Meiosis
in here?
Sexual reproduction occurs through the production of gametes which
contain only one set of genes (haploid, 1N). When two gametes fuse, a
new diploid organism with a different complement of gene copies than
either parent is produced. Haploid gametes are produced through the
process of meiosis.
Figure 8:
Cell cycle subdivisions and
check points
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(continued on next page)
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Background (COntinued)
Notes
Meiosis consists of one round of DNA replication followed by
two nuclear divisions, meiosis I and meiosis II. This results in the
formation of four daughter cells, each with only half the number
of chromosomes of the parent. When two gametes combine during
fertilization to form a zygote, the diploid chromosome number is
restored in the resulting organism. Typically, we think of gametes as
cells that come from either a male (sperm) or a female (ova) that are
specialized to fuse with each other in species specific way (mouse
sperm cannot fertilize a whale ovum).
Under a microscope, mitosis and meiosis do not look very different.
The phases of division are visually distinct and named as their visual
look-alikes in mitosis. However, the way replicated DNA is divided is
very different in meiosis than mitosis.
Prophase I:
The duplicated sister chromosomes condense and are attached at
a centromere as in mitosis. Unlike mitosis, these sister pairs form
a tetrad with duplicated chromosome homologs from both the
maternally and paternally contributed chromosomes. At this point
DNA may cross over between non-sister, chromatid homologs
forming unique chromatids that contain DNA from both maternal
and paternal chromosomes.
Metaphase I:
The tetrads align at the center of the cell and spindle microtubules
attach to the kinetochores.
Anaphase I:
The tetrads separate into two, duplicate chromosomes that remain
attached at the centromere which move to separate poles of the
cell.
Anaphase I is followed by telophase 1 and cytokinesis, resulting in
two daughter cells that each have 2N DNA.
Mitosis II follows immediately after meiosis I is completed without
further duplication of the DNA. The single set of chromosomes (held
together at the centromere) are separated on the second spindle thus
forming daughter cells that are now 1N. A starting diploid germ cell
has now formed 4 haploid gametes, each containing a mixture of DNA
(continued on next page)
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Background (COntinued)
Notes
from both the mother and the father. This method of division produces
gametes that vary greatly between each other with respect to the allele
combinations available to pass to the next generation.
Figure 9 (reprinted from College Board)
Crossover events can be observed in fungal asci resulting
from meiotic divisions
The example of meiosis
that will be used in this
investigation is Sordaria
fimicola. S. fimicola is an
ascomycete fungus that
is haploid for the bulk of
its life cycle; the haploids
comprise the individual
fungal filaments, called
hyphae, which normally
exist in a mass, called a
mycelium, representing
the “body” of the fungus,
and the ascospores, from
which mycelia develop.
The only diploid portion of
the life cycle of S. fimicola
occurs when the nuclei of
specialized hyphae come
together.
Figure 10: Life cycle of the fungus,
Sordaria.
(continued on next page)
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Background (COntinued)
Notes
These hyphae fuse to form a diploid zygote. This zygote then
undergoes meiosis to produce the haploid ascospores, yielding four
haploid nuclei contained in a sac called an ascus. After meiosis I
and II, the four haploid nuclei undergo mitosis, resulting in an ascus
containing eight haploid ascospores from one diploid zygote. Many
asci form inside a fruiting body called a perithecium (Figure_5_).
Wild types sordaria are a dark brown, but we will grow this culture
in the presence of a tan mutant so that when mycelium from a wild
type fuses with the tan to form a zygote, we will be able to observe the
results of crossing over events that occurred as the zygote underwent
meiotic division. Figure 6 shows a culture plate that was seeded with
two squares of wild type sordaria containing agar and two squares of
tan type sordaria containing agar. The mycelium grow out from these
blocks so fusion will occur where the two types meet. This region of
fusion is where you will find perithecium containing ascospores from
zygotes that came from a wild type and a tan parent (arrows). After
meiosis I and II, the haploid ascospores will be either tan or wild type.
When the parathecium are gently ruptured, the ascospores will be in
strings of 8 that can easily be scored as tan or wild.
Figure 11
Figure 12???
Insert SORDIA Image on
Page 7 250-7052???
Figure 12: Meisosis with no crossing over
(continued on next page)
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Background (COntinued)
Notes
How these ascospores are arranged within the ascus is a direct representation of
whether or not crossing over has occurred between the centromere and the site for
the gene for ascospore color. If no crossing over has occurred, the ascospores will
be arranged in a 4:4 manner. If crossing over has occurred, they will occur in a
2:4:2 or 2:2:2:2 manner(s).
Figure 13: Meisosis with crossing over
By observing the ascospore arrangement, the percentage of asci exhibiting
crossover can be determined. This frequency appears to be affected largely by
the distance from the gene to the centromere. From the crossover frequency, the
distance in map units from the gene for ascospore color, and the chromosome
centromere, can be calculated.
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Notes
Kit # 3674-07
Safety Precautions
‹ When working with the chromatography solvent, use a
chemical hood or proper ventilation.
‹ As general safe laboratory practice, it is recommended that you
wear proper protective equipment, such as gloves, safety goggles,
and a lab apron.
‹ As general lab practice, read the lab through completely before
starting, including any Material Safety Data Sheets (MSDSs) and
live materials care sheets at the end of this booklet as well as any
appropriate MSDSs for any additional substances you would like
to test. One of the best sources is the vendor for the material. For
example, when purchased at Wards, searching for the chemical on
the Ward’s website will direct you to a link for the MSDS. (Note:
There are no live materials care sheets included in this particular
lab.)
At the end of the lab:
‹ All laboratory bench tops should be wiped down with a 20%
bleach solution or disinfectant to ensure cleanliness.
‹ Wash your hands thoroughly with soap and water before leaving
the laboratory.
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Procedure
Tips
Kit # 3674-07
Part 1 – Structured INQUIRY
MATERIALS needed PER LAB GROUP
‹ When performing this lab
activity, all data should be
recorded in a lab notebook. You
will need to construct your own
data tables, where appropriate,
in order to accurately capture
the data from the investigation.
q
q
q
q
‹ If directed to do so by your
teacher, this part of the lab may
be done at the same time as Part
2 of the lab.
Part 1a – PROCEDURE: modeling mitosis
???
???
??
??
Red pop beads
Pink pop beads
‹
Clear plastic cetrioles
Magnetic yellow centromeres
please fix/edi list for this all parts of
this activity or remove completely
Using the pop beads (red and pink), the magnetic yellow centomeres,
and the clear plastic centrioles, simulate each stage of mitosis.
To simulate Interphase – DNA replication:
1. Construct two strands of seven red pop beads and attach each
strand to a yellow centromere. Repeat with two strands of seven
pink pop beads and a yellow centromere. These will represent a
homologous pair of chromosomes (red from the father and pink
from the mother).
2. Picture an imaginary boundary in the center of your desk.
This boundary will represent the nuclear membrane. Place the
chromosomes in the center of the imaginary nucleus.
3. DNA replication occurs, producing a duplicate of each
chromosome. Construct two chromosomes identical to the ones
you made previously. Each of the duplicated chromosomes is
called a chromatid. Join both red chromatids at the centromere
to form a pair of sister chromatids. Repeat this process for the
pink chromosome.
4. Place a pair of plastic centrioles, at a ninety degree angle, just
outside of your nuclear membrane. The centrioles also replicated
during interphase, so place another pair next to them in your
cell.
‹
TIP: It may be helpful to tape your centrioles together
during the exercise.
5. Illustrate your simulation of interphase and in your own words
provide a brief description of what happens during interphase.
(continued on next page)
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
PROCEDURE – Part 1a: modeling mitosis (continued)
Notes
To simulate Prophase:
1. Move your two pairs of centrioles to opposite poles (sides) of
the cell (your desk).
2. Illustrate your simulation of prophase and in your own words
provide a brief description of what happens during prophase.
To simulate Metaphase:
1. Center your chromosomes along an imaginary metaphase plate
with the centrioles still at the opposite poles of the cells.
2. Illustrate your simulation of metaphase and in your own words
provide a brief description of what happens during metaphase.
To simulate Anaphase:
1. Separate and move the centromeres of each chromosome
toward opposite poles of the cell. Notice how the arm of each
chromosome trails the centromeres to the poles.
2. Illustrate your simulation of anaphase and in your own words
provide a brief description of what happens during anaphase.
To simulate Telophase and Cytokinesis:
1. Move one red strand and one pink strand to the centrioles they
were heading toward during anaphase. Imagine a cleavage furrow
developing between each nuclei and separating the cell into two
daughter cells.
2. Note how each cell now contains one red and one pink
chromosome, as well as one pair of centrioles, exactly like the cell
with which you began.
3. Illustrate your simulation of telophase/cytokinesis and in your
own words provide a brief description of what happens during
telophase/cytokinesis.
(continued on next page)
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Notes
Kit # 3674-07
Procedure– Part 1b:
Observing Mitosis in Animal and Plant Cells
1. Obtain a compound microscope, the fish blastula slide and the
onion mitosis slide.
2. Observe the prepared microscope slide of onion root tip first
at 100X (10X objective and 10X ocular) then at 400X (40X
objective, 10X ocular). Look for cells in mitosis.
‹
Depending on the quality of your microscope you may be
able to distinguish the various phases of mitosis.
‹
Use oil immersion if available.
Using the mitosis illustration provided on page ___ as a guide,
try to identify each phase of animal cell mitosis.
‹ blastodisc???
3. Observe the prepared microscope slide of the fish blastodisc
mitosis first at 100X, then 400X. Look for cells in mitosis and
classify the stages. Compare and record similarities and contrast
differences between the animal and plant cell mitosis.
4. Examine at least three fields of view of the apical meristem of
the onion root tip at 400X. In each view, count and record the
number of cells in interphase and the various stages of mitosis.
5. Calculate the percentage of total cells counted in interphase
and in each stage of mitosis. If the individual phases cannot be
discerned, calculate the percentage in interphase and in mitosis.
Table 1
# cells
field 1
Stage
# cells
field 2
# cells
field 3
Total #
cells
% total
number
of cells
Time
in each
stage
Interphase
Prophase
Metaphase
Anaphase
Telophase
(continued on next page)
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Notes
Kit # 3674-07
Procedure– Part 1b: Observing Mitosis in
Animal and Plant Cells (continued)
6. Assuming that it takes an average of 24 hours (1,440 minutes)
for onion root tip cells to complete the cell cycle, calculate the
amount of time cells spent in each phase of the cycle. Use the
formula provided below. Enter your results in Table 1.
% of cells in phase x 1,440 minutes = _______ minutes cell spent in phase
7. Make a pie chart representing the amount of time spent in each
stage of mitosis.
‹
Prophase is normally the longest phase of mitosis, due to
the complexity of the events occurring during prophase.
These complex events take a relatively long time for the
cell to perform:chromatin condensing and thickening,
nuclear membrane dissolving, and the early stages of
spindle development.
Procedure– Part 1C: garlic root tip squash
Due to time constraints, your instructor may have grown garlic root
tips in advance. If this is the case, begin with Step 8.
1. Obtain a jar or Petri dish.
2. Using a ruler, measure 1.5 cm starting at the bottom base of the
jar and mark the measurement with a permanent marker.
3. Fill the jar with the fine sand to the mark on the jar. You may
need to sift or swirl the sand back and forth to ensure you have
distributed the sand evenly.
4. Add water to the jar to wet the sand.
5. Separate a clove of garlic into individual sections, or “toes.”
Remove the paper-like skin from each section and any dried
roots on the primordia (blunt end) with a razor blade.
‹
The section used must have root primordia present or it
will not produce root tips. The root tips will grow within
two days for very fresh garlic, or three days for older
garlic. Because garlic has a mitotic cycle of approximately
12.5 hours, it is important to “plant” the garlic and harvest
the root tips at approximately the same time of day in
order to get the greatest percentage of meristematic cells
undergoing mitosis.
(continued on next page)
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Procedure– Part 1C: garlic root tip squash
Notes
(continued)
6. Place the cloves into the jar with the root primordia (blunt end)
in the sand.
7. Place the jar in a box or dark place until the root tips have grown
to a length of about 4 to 5 mm. Several viable root tips will
grow on each section of garlic.
‹
It is important that the garlic grows in the dark to ensure that
it produces roots rather than shoots.
‹
Remove the garlic from the box approximately one halfhour before performing the experiment to expose the root
tips to light.
8. Blot as much excess water from the root tips as possible. Any
excess water on the slide will affect your results. Do not allow
the root tips to dry out, however.
9. Using a scalpel, cut off the end of one of the emergent root tips;
the section should be approximately 1 to 2 mm long. Place the
root tip on a clean microscope slide and apply two or three drops
of hydrochloric acid (HCl) to the root tip.
10. Holding the slide with a clothespin, pass it through the flame of
a Bunsen burner for five seconds.
‹
Pass the slide through the flame of the Bunsen burner. Do
not hold the slide directly in or over the flame.
11. Without harming the root tip, blot the specimen with a paper
towel to remove the excess HCl.
‹
You may wish to touch a corner of the paper towel to the
drop on the slide and allow the paper towel to soak it up.
This may not remove the liquid from the slide as effectively
as blotting, but it will not disturb the root tip.
12. Add a few drops of carbol fuchsin stain, covering the root tip.
13. Pass the slide through the flame of the Bunsen burner for two
minutes. Let the slide stand for one minute.
‹
Pass the slide through the flame of the Bunsen burner. Do
not hold the slide directly in or over the flame.
(continued on next page)
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Procedure– Part 1C: garlic root tip squash
Notes
(continued)
14. Without disturbing the specimen, use a paper towel to remove
the excess stain.
15. Cover with a coverslip. Using a pencil eraser or other blunt
instrument, gently press down on the coverslip to squash and
spread out the root tip. Blot off the excess stain, if any, that may
have come out from under the coverslip.
16. Observe the slide under a microscope at 100X. Locate the apical
meristem. Examine the slide at 400X. Locate cells in the various
stages of mitosis, and make sketches of what you find.
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Procedure
TipS
‹ If directed to do so by your
teacher, this part of the lab may
be done at the same time as Part
1a of the lab.
Kit # 3674-07
Part 2 – GUIDED INQUIRY: MEIOSIS
MATERIALS needed PER LAB GROUP
q PLEASE LIST FOR THIS PROCEDURE OR REMOVE
COMPLETELY
Part 2A – PROCEDURE: modeling meiosis
1. Using the colored pop beads follow a similar procedure as you
did for mitosis, but model meiosis I and meiosis II.
Part 2b – PROCEDURE:
Meiosis and Sordaria Crossing Over
1. Place a drop of water on each of the clean microscope slides
with an inoculating loop. Within this procedure, you will
prepare three slides to get an adequate sampling of hybrids.
2. With an inoculating loop, scrape several perithecia from the
demo cross plate. Scrape the perithecia from the interface of the
two crossing strains (Figure _6_) close to the edge of the plate
and place in the drop of water on the slide. Avoid picking up
agar along with perithecia; it will interfere with the results.
3. Cover the slide with a coverslip. Using a pencil eraser or other
blunt instrument, gently press down on the coverslip to squash
and spread out the perithecia. The pressure should be sufficient
to squeeze asci from the perithecia, but not enough to crush the
asci themselves.
Figure __: Sordaria squash
Observe the slide under a microscope at 100X. Locate the
asci. Then view the slide at 400X to determine the color of
the ascospores. The slide preparation should show collapsed
perithecia and asci clusters (rosettes), with mature ascospores
in various arrangements. Immature ascospores will all be light
colored. Since Sordaria fimicola is homothallic, the preparation
will show both hybrid and self-fertilized perithecia, however,
fertilization will not occur very far from the line of contact
between the two varieties.
4. Count approximately 50 hybrid asci from at least three fields of
view, preferably from different slides. Record the data in your
laboratory sheet or notebook.
(continued on next page)
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Notes
Kit # 3674-07
Part 2b – PROCEDURE:
Meiosis and Sordaria Crossing Over (continued)
5. Calculate the frequency (%) of asci crossing-over.
% Cross over = # showing cross over x 100
total counted
‹
TIP: The percentage of crossing-over must be divided by
two, since only half the ascospores in each hybrid ascus are
the result of crossing over.
6. Determine the number of map units between the centromere and
the gene for ascospore color.
7. Compare your data with the class and make an account for the
classrooms data versus published data about sordaria crossing
over. The published distance between the tan gene and the
centromere is 26 map units.
8.
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Dispose of lab materials.
‹
NOTE: Properly dispose of sharps.
‹
When finished with the sordaria plate, please dispose of it
in one of the following ways:
•
Use a 20% bleach solution for 10 minutes.
•
Place the organism in 70% isopropanol alcohol for
24 hours.
•
Autoclave the organism at 121 °C for 15 minutes in an
autoclave bag. The Petri dish will melt in the autoclave
so be sure to bag your organism and close securely
before autoclaving.
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Part 2 assessment questions
1. How does meiosis lead to genetic variability within a population? Use S. fimicola as an example.
Meiosis leads to genetic variability through the segregation of gene alleles, the independent assortment
of genes, and crossing-over, as well as the variability that results from the combination of the genetic
material from the gametes of two genetically different individuals.
2. How does genetic variability represent an adaptive advantage for organisms that reproduce sexually?
With increased variability among individuals in a population comes an increased probability that the
population would be able to survive under changing environmental and evolutionary pressures. This
ability, in turn, gives the species a better chance of surviving and thriving in the future.
3. Why is S. fimicola an ideal organism for the demonstration of crossing-over?
The fact that it displays both haploid and diploid stages of reproduction allows scientists to easily
manipulate different strains of the organism. The colored ascospores are easily identified and the
ascospore patterns readily indicate when crossing-over has occurred.
4. Research cell division in prokaryotic and eukaryotic cells, compare and contrast the characteristics of
mitosis in each.
Unlike eukaryotic cells, prokaryotic cells utilize neither mitosis nor meiosis. Prokaryotic cells replicate
through a process known as binary fission. The prokaryotic DNA, which is found free in the cell and
not confined in a membrane-bound nucleus, replicates and each copy attaches to a different part of
the cell’s plasma membrane. The cell grows and, when it is approximately double its original size, the
membrane grows inward, dividing the cell into two genetically identical daughter cells.
5. Using what you know about mitosis, genetic transcription, cell cycle, and the two karyotype images
answer the following questions:
How do cells ensure DNA replication is accurate?
The DNA polymerase has a 3’---5’ exonucluease domain. This domain is responsible for proofreading
the DNA and reads in the opposite direction of DNA replication.
According to the images, on which chromosomes did the mutation occur? By which mechanism of cell
replication do you think the Philadelphia chromosome occurs? Explain.
????
Assuming that it takes an average of 12 hours for CML cells to complete the cell cycle, how many cells
will be produced within a week (7days)?
4,096 cells will develop in one week (168 hours/week)
Would you agree that cancer is the disease of mitosis? Explain.
Answers will vary, but the students should be strongly considering cancer as a disease of mitosis.
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
EXPERIMENT
DESIGN Tips
The College Board encourages peer
review of student investigations
through both formal and informal
presentation with feedback and
discussion. Assessment questions
similar to those on the AP exam
might resemble the following questions, which also might arise in peer
review:
‹ Explain the purpose of a
procedural step.
‹ Identify the independent
variables and the dependent
variables in an experiment.
‹ What results would you expect
to see in the control group? The
experimental group?
‹ How does XXXX concept
account for YYYY findings?
‹ Describe a method to determine
XXXX.
Kit # 3674-07
Part 3: Cell Processes: Mitosis and Meiosis
open inquiry: design an experiment
What questions occurred to you as you investigated mitosis and
meiosis? Now that you are familiar with a mitotic system (onion root
tip squash) and a meiotic system (Sordaria ascus formation using a tan
mutant) design an experiment to investigate one of your questions.
Questions may involve differences in mitotic rates in different cell
types, what types of environmental signals would inhibit or accelerate
mitosis, what types of environmental conditions would result in the deregulation of mitosis or meiosis, are there conditions that would inhibit
or accelerate crossing over during meiosis, how might mitosis proceed
differently in a cancerous cell line, how are germ cells that produce
haploid daughter cells different from somatic cells that produce diploid
cells?
See optional materials not provided for some agents that affect mitotic
rate in some systems.
Before starting your experiment, plan your investigation in your lab
notebook. Have your teacher check over and initial your experiment
design. Once your design is approved, investigate your hypothesis.
Be sure to record all observations and data in your laboratory sheet or
notebook.
Use the following steps when designing your experiment.
1. Define the question or testable hypothesis.
2. Describe the background information. Include previous
experiments.
3. Describe the experimental design with controls, variables, and
observations.
4. Describe the possible results and how they would be interpreted.
5. List the materials and methods to be used.
6. Note potential safety issues.
After the plan is approved by your teacher:
7. The step by step procedure should be documented in the
lab notebook. This includes recording the calculations of
(continued on next page)
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Part 3: open inquiry (continued)
Notes
concentrations, etc. as well as the weights and volumes used.
8. The results should be recorded (including drawings, photos, data
print outs).
9. The analysis of results should be recorded.
10. Draw conclusions based on how the results compared to the
predictions.
11. Limitations of the conclusions should be discussed, including
thoughts about improving the experimental design, statistical
significance and uncontrolled variables.
12. Further study direction should be considered.
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Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
EXTENSION ACTIVITY: Loss of cell cycle control
Notes
NOTE: Cancer was chosen as the example for this exercise because it
demonstrates what occurs when a cell loses control of the cell cycle.
There is a possibility that your student will have a family member with
cancer, or who died of cancer. It may be appropriate to suggest that
they discuss the issue with their family or seek the advice of a medical
practitioner.
Procedure
When performing this lab activity, all data should be recorded in a
legal scientific notebook or on the laboratory sheet provided. Students
will need to construct their own data tables, where appropriate, in
order to neatly and accurately capture the data from the lab activity
and their investigations.
Leukemia is a type of cancer that is characterized by the uncontrolled
growth of a specific type of leukocyte (white blood cell). The
Philadelphia chromosome causes several kinds of leukemia; for
example, Chronic Myelogenous Leukemia (CML). During this part of
the lab, compare karyotypes between normal and CML.
Search the Internet for images of normal and CML karyotypes. Be sure
to print the image as it will be needed for the next step.
Compare and contrast between a normal chromosomes vs. CML
chromosomes.
Note. Be sure both karyotypes are from the same sex, either both male
or both female.
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Page 31
Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Material safety data sheets
Material Safety Data Sheet
MSDS # 164.00
Section 1:
Page 1 of 2
Carbol Fuchsin Stain
Product and Company Identification
Carbol Fuchsin Stain
Synonyms/General Names: Carbol Fuchsin, Biological Stain
Product Use: For educational use only
Manufacturer: Columbus Chemical Industries, Inc., Columbus, WI 53925.
24 Hour Emergency Information Telephone Numbers
CHEMTREC (USA): 800-424-9300
CANUTEC (Canada): 613-424-6666
ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com
Section 2:
Hazards Identification
Opaque red liquid, characteristic phenol odor
WARNING! Moderately toxic by ingestion and severely corrosive to body tissue.
Target organs: Central nervous system, liver, kidneys, eyes.
HMIS (0 to 4)
Health
2
Fire Hazard
0
Reactivity 0
This material is considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200).
Section 3:
Composition / Information on Ingredients
Ethyl Alcohol, (64-17-5), 6-7%.
Fuchsin Basic, (632-99-5), <1%.
Phenol, 90%, Liquified, (108-95-2), 4-5%.
Water, (7732-18-5), 87-88%.
Section 4:
Eyes:
Skin:
Ingestion:
Inhalation:
First Aid Measures
Always seek professional medical attention after first aid measures are provided.
Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.
Immediately flush skin with excess water for 15 minutes while removing contaminated clothing.
Call Poison Control immediately. Rinse mouth with cold water. Give victim 1-2 cups of water or milk to drink.
Induce vomiting immediately.
Remove to fresh air. If not breathing, give artificial respiration.
Section 5:
Fire Fighting Measures
When heated to decomposition, emits acrid fumes.
Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire.
Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool
container with water spray. Material is not sensitive to mechanical impact or static discharge.
Section 6:
2
0
0
Accidental Release Measures
Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected
personnel. Remove all ignition sources and ventilate area. Contain spill with sand or absorbent material and place material in a
sealed bag or container for disposal. Wash spill area after pickup is complete. See Section 13 for disposal information.
Section 7:
Handling and Storage
Green
Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash
hands thoroughly after handling.
Storage: Store in General Storage Area [Green Storage] with other items with no specific storage hazards. Store in a cool, dry,
well-ventilated, locked store room away from incompatible materials.
Section 8:
Exposure Controls / Personal Protection
Use ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire
extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash
hands thoroughly after handling material and before eating or drinking. Exposure guidelines: Ethyl Alcohol: OSHA PEL: 1900
mg/m3 and ACGIH TLV: 1000 ppm, STEL: N/A. Phenol: OSHA PEL: 19 mg/m3, ACGIH: TLV: 19 mg/m3, STEL: 60 mg/m3
ceiling/skin. Fuchsin Basic Stain: OSHA PEL: N/A, ACGIH: TLV: N/A, STEL: N/A
© 2008, Scholar Chemistry. All Rights Reserved.
1/12/2012
(continued on next page)
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
US: www.wardsci.com
Canada: www.wardsci.ca
250-7456 v.5/12
Page 32
Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Material safety data sheets
Material Safety Data Sheet
MSDS # 164.00
Section 9:
Molecular formula
Molecular weight
Specific Gravity
Vapor Density (air=1)
Melting Point
Boiling Point/Range
Vapor Pressure (20°C)
Flash Point:
Autoignition Temp.:
Page 2 of 2
Carbol Fuchsin Stain
Scholar Chemistry
Physical and Chemical Properties
Mixture.
N/A.
0.9846 g/mL @ 20°C.
0.7 (water).
0°C.
100°C.
N/A.
118ºC (244 ºF) CC Ethanol.
N/A.
Section 10:
Appearance
Odor
Odor Threshold
Solubility
Evaporation rate
Partition Coefficient
pH
LEL
UEL
Opaque red liquid..
Characteristic phenol odor.
N/A.
Complete.
N/A (Butyl acetate = 1).
N/A (log POW).
N/A.
4.0% Ethanol.
20.0% Ethanol.
N/A = Not available or applicable
Stability and Reactivity
Stability: Stable under normal conditions of use and storage.
Incompatibility: Oxidizing agents, acids, halogens, calcium hypochlorite.
Shelf life: Indefinite if stored properly.
Section 11:
Toxicology Information
Acute Symptoms/Signs of exposure: Eyes: Redness, tearing, itching, burning, conjunctivitis. Skin: Redness, itching.
Ingestion: Irritation and burning sensations of mouth and throat, nausea, vomiting and abdominal pain. Inhalation: Irritation of
mucous membranes, coughing, wheezing, shortness of breath,
Chronic Effects: No information found.
Sensitization: none expected
Ethyl Alcohol: LD50 [oral, rat]; 7060 mg/kg; LC50 [rat]; 20,000 mg/l (10 hours); LD50 Dermal [rabbit]; 20 mg/24H MOD
Phenol: LD50 [oral, rat]; 317 mg/kg; LC50 [rat]; 316 mg/m3; LD50 Dermal [rabbit]; 500mg/24 hrs/severe.
Fuchsin Basic Stain: LD50 [oral, rat]; N/A; LC50 [rat]; N/A; LD50 Dermal [rabbit]; N/A
Material has not been found to be a carcinogen nor produce genetic, reproductive, or developmental effects.
Section 12:
Ecological Information
Ecotoxicity (aquatic and terrestrial):
environment
Toxic to terrestrial and aquatic plants and animals. Do not release to the
Section 13:
Disposal Considerations
Check with all applicable local, regional, and national laws and regulations. Local regulations may be more stringent than
regional or national regulations. Small amounts of this material may be suitable for sanitary sewer or trash disposal.
Section 14:
DOT Shipping Name:
DOT Hazard Class:
Identification Number:
Transport Information
Not regulated by DOT.
Section 15:
EINECS: Not listed .
TSCA: All components are listed or are exempt.
Canada TDG: Not regulated by TDG.
Hazard Class:
UN Number:
Regulatory Information
WHMIS Canada: Not WHMIS Controlled.
California Proposition 65: Not listed.
The product has been classified in accordance with the hazard criteria of the Controlled Products Regulations and the MSDS
contains all the information required by the Controlled Products Regulations.
Section 16:
Other Information
Current Issue Date: January 12, 2012
Disclaimer: Scholar Chemistry and Columbus Chemical Industries, Inc., (“S&C”) believes that the information herein is factual but is not intended to be all
inclusive. The information relates only to the specific material designated and does not relate to its use in combination with other materials or its use as to any
particular process. Because safety standards and regulations are subject to change and because S&C has no continuing control over the material, those
handling, storing or using the material should satisfy themselves that they have current information regarding the particular way the material is handled, stored
or used and that the same is done in accordance with federal, state and local law. S&C makes no warranty, expressed or implied, including (without
limitation) warranties with respect to the completeness or continuing accuracy of the information contained herein or with respect to fitness for any
particular use.
© 2008, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
1/12/2012
US: www.wardsci.com
Canada: www.wardsci.ca
250-7456 v.5/12
Page 33
Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Material safety data sheets
Material Safety Data Sheet
Hydrochloric Acid Solution, 1.0M
MSDS # 338.00
Section 1:
Page 1 of 2
Product and Company Identification
Hydrochloric Acid Solution, 1.0M
Synonyms/General Names: Muriatic Acid; Hydrochloric Acid Solution, 1N
Product Use: For educational use only
Manufacturer: Columbus Chemical Industries, Inc., Columbus, WI 53925.
24 Hour Emergency Information Telephone Numbers
CHEMTREC (USA): 800-424-9300
CANUTEC (Canada): 613-424-6666
ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com
Section 2:
Hazards Identification
Clear colorless liquid; pungent odor.
WARNING! Strongly corrosive to body tissue and moderately toxic by ingestion.
Target organs: Respiratory system, eyes, skin, lungs.
HMIS (0 to 4)
Health
2
Fire Hazard
0
Reactivity 0
This material is considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200).
Section 3:
Composition / Information on Ingredients
Hydrochloric Acid, 37% (7647-01-0), 9-10%.
Section 4:
Eyes:
Skin:
Ingestion:
Inhalation:
Water (7732-18-5), 90-91%.
First Aid Measures
Always seek professional medical attention after first aid measures are provided.
Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.
Immediately flush skin with excess water for 15 minutes while removing contaminated clothing.
Call Poison Control immediately. Do not induce vomiting. Rinse mouth with cold water. Give victim 1-2 cups of
water or milk to drink.
Remove to fresh air. If not breathing, give artificial respiration.
Section 5:
Fire Fighting Measures
When heated to decomposition, emits acrid fumes.
Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire.
Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool
container with water spray. Material is not sensitive to mechanical impact or static discharge.
Section 6:
2
0
0
Accidental Release Measures
Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected
personnel. Remove all ignition sources and ventilate area. Contain spill with sand or absorbent material and place material in a
sealed bag or container for disposal. Wash spill area after pickup is complete. See Section 13 for disposal information.
Section 7:
Handling and Storage
White
Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash
hands thoroughly after handling.
Storage: Store in Corrosive Area [White Storage] with other corrosive items. Store in a dedicated corrosive cabinet. Store in a
cool, dry, well-ventilated, locked store room away from incompatible materials.
Section 8:
Exposure Controls / Personal Protection
Use ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire
extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash
hands thoroughly after handling material and before eating or drinking. Use NIOSH-approved respirator with an acid/organic
cartridge. Exposure guidelines Hydrochloric Acid: OSHA PEL: 5 ppm ceiling and ACGIH TLV: 2 ppm ceiling, STEL: N/A.
© 2008, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
(continued on next page)
US: www.wardsci.com
Canada: www.wardsci.ca
12/20/2011
250-7456 v.5/12
Page 34
Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Material safety data sheets
Material Safety Data Sheet
MSDS # 338.00
Section 9:
Molecular formula
Molecular weight
Specific Gravity
Vapor Density (air=1)
Melting Point
Boiling Point/Range
Vapor Pressure (20°C)
Flash Point:
Autoignition Temp.:
Page 2 of 2
Hydrochloric Acid, 1.0M
Scholar Chemistry
Physical and Chemical Properties
HCl.
36.46.
1.01 g/mL @ 20°C.
0.7.
0°C.
100°C.
14.
N/A.
N/A.
Section 10:
Appearance
Odor
Odor Threshold
Solubility
Evaporation rate
Partition Coefficient
pH
LEL
UEL
Clear, colorless liquid.
Pungent odor.
N/A.
Completely soluble in water.
<1
(Butyl acetate = 1).
N/A.
(log POW).
1, acid (corrosive).
N/A.
N/A.
Stability and Reactivity
Avoid heat and ignition sources.
Stability: Stable under normal conditions of use and storage.
Incompatibility: Alkalis, strong bases, metals, amines, carbonates, metal oxides, cyanides, sulfides, sulfites and formaldehyde.
Shelf life: Indefinite, store in a cool, dry environment.
Section 11:
Toxicology Information
Acute Symptoms/Signs of exposure: Eyes: Redness, tearing, itching, burning, damage to cornea, conjunctivitis, loss of vision.
Skin: Redness, blistering, burning, itching, tissue destruction with slow healing. Ingestion: Nausea, vomiting, burning, diarrhea,
ulceration, convulsions, shock. Inhalation: Coughing, wheezing, shortness of breath, headache, spasm, inflammation and edema
of bronchi, pneumonitis.
Chronic Effects: Repeated/prolonged skin contact may cause thickening, blackening or cracking. Repeated eye exposure may
cause corneal erosion or loss of vision.
Sensitization: none expected
Hydrochloric Acid: LD50 [oral, rabbit]; 900 mg/kg; LC50 [rat]; 3124 ppm (1 hour); LD50 Dermal [rabbit]; N/A
Material has not been found to be a carcinogen nor produce genetic, reproductive, or developmental effects.
Section 12:
Ecological Information
Ecotoxicity (aquatic and terrestrial):
LC50 - 282 mg/l - 96 h - Gambusia affinis (Mosquito fish)
Section 13:
Disposal Considerations
Check with all applicable local, regional, and national laws and regulations. Local regulations may be more stringent than
regional or national regulations. Small amounts of this material may be suitable for sanitary sewer disposal after being
neutralized to pH 7.
Section 14:
DOT Shipping Name:
DOT Hazard Class:
Identification Number:
Transport Information
Hydrochloric Acid.
8, pg II .
UN1789.
Section 15:
Canada TDG:
Hazard Class:
UN Number:
Hydrochloric Acid.
8, pg II.
UN1789.
Regulatory Information
EINECS: Listed (231-595-7).
WHMIS Canada: CLASS E: Corrosive liquid.
TSCA: All components are listed or are exempt.
California Proposition 65: Not listed.
The product has been classified in accordance with the hazard criteria of the Controlled Products Regulations and the MSDS
contains all the information required by the Controlled Products Regulations.
Section 16:
Other Information
Current Issue Date: December 20, 2011
Disclaimer: Scholar Chemistry and Columbus Chemical Industries, Inc., (“S&C”) believes that the information herein is factual but is not intended to be all
inclusive. The information relates only to the specific material designated and does not relate to its use in combination with other materials or its use as to any
particular process. Because safety standards and regulations are subject to change and because S&C has no continuing control over the material, those
handling, storing or using the material should satisfy themselves that they have current information regarding the particular way the material is handled, stored
or used and that the same is done in accordance with federal, state and local law. S&C makes no warranty, expressed or implied, including (without
limitation) warranties with respect to the completeness or continuing accuracy of the information contained herein or with respect to fitness for any
particular use.
© 2008, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
(continued on next page)
US: www.wardsci.com
Canada: www.wardsci.ca
12/20/2011
250-7456 v.5/12
Page 35
Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Material safety data sheets
Material Safety Data Sheet
Isopropyl Alcohol, 70%
MSDS # 384.00
Section 1:
Page 1 of 2
Product and Company Identification
Isopropyl Alcohol, 70%
Synonyms/General Names: N/A
Product Use: For educational use only
Manufacturer: Columbus Chemical Industries, Inc., Columbus, WI 53925.
24 Hour Emergency Information Telephone Numbers
CHEMTREC (USA): 800-424-9300
CANUTEC (Canada): 613-424-6666
ScholAR Chemistry; 5100 W. Henrietta Rd, Rochester, NY 14586; (866) 260-0501; www.Scholarchemistry.com
Section 2:
Hazards Identification
HMIS (0 to 4)
Health
1
Fire Hazard
3
Reactivity 0
Clear, colorless liquid, alcohol odor.
WARNING! Flammable liquid and slightly toxic by ingestion.
Flammable liquid, keep away from all ignition sources.
Target organs: Central nervous system, liver, kidneys.
This material is considered hazardous by the OSHA Hazard Communication Standard (29 CFR 1910.1200).
Section 3:
Composition / Information on Ingredients
Isopropyl Alcohol (67-63-0), 64%.
Water (7732-18-5), 36%.
Section 4:
Eyes:
Skin:
Ingestion:
Inhalation:
First Aid Measures
Always seek professional medical attention after first aid measures are provided.
Immediately flush eyes with excess water for 15 minutes, lifting lower and upper eyelids occasionally.
Immediately flush skin with excess water for 15 minutes while removing contaminated clothing.
Call Poison Control immediately. Aspiration hazard. Rinse mouth with cold water. Give victim 1-2 tbsp of
activated charcoal mixed with 8 oz water.
Remove to fresh air. If not breathing, give artificial respiration.
Section 5:
Fire Fighting Measures
Class IB Flammable Liquid. When heated to decomposition, emits acrid fumes
Protective equipment and precautions for firefighters: Use foam or dry chemical to extinguish fire.
Firefighters should wear full fire fighting turn-out gear and respiratory protection (SCBA). Cool container
with water spray. Material is not sensitive to mechanical impact. Material is sensitive to static discharge.
Section 6:
1
3
0
Accidental Release Measures
Use personal protection recommended in Section 8. Isolate the hazard area and deny entry to unnecessary and unprotected
personnel. Remove all ignition sources and ventilate area. Contain spill with sand or absorbent material and place material in a
sealed bag or container for disposal. Wash spill area after pickup is complete. See Section 13 for disposal information.
Section 7:
Handling and Storage
Red
Handling: Use with adequate ventilation and do not breathe dust or vapor. Avoid contact with skin, eyes, or clothing. Wash
hands thoroughly after handling.
Storage: Store in Flammable Area [Red Storage] with other flammable materials and away from any strong oxidizers. Store in a
dedicated flammables cabinet. Store in a cool, dry, well-ventilated, locked store room away from incompatible materials.
Section 8:
Exposure Controls / Personal Protection
Use ventilation to keep airborne concentrations below exposure limits. Have approved eyewash facility, safety shower, and fire
extinguishers readily available. Wear chemical splash goggles and chemical resistant clothing such as gloves and aprons. Wash
hands thoroughly after handling material and before eating or drinking. Use NIOSH-approved respirator with an acid/organic
cartridge. Exposure guidelines: Isopropyl Alcohol: OSHA PEL: 980 mg/m3, ACGIH TLV: 492 mg/m3, STEL: 984 mg/m3.
© 2008, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
12/21/2011
US: www.wardsci.com
Canada: www.wardsci.ca
250-7456 v.5/12
Page 36
Cell Processes: Mitosis and Meiosis – Teacher’s Guide
Kit # 3674-07
Material safety data sheets
Material Safety Data Sheet
MSDS # 384.00
Section 9:
Molecular formula
Molecular weight
Specific Gravity
Vapor Density (air=1)
Melting Point
Boiling Point/Range
Vapor Pressure (20°C)
Flash Point:
Autoignition Temp.:
Page 2 of 2
Isopropyl Alcohol, 70%
Scholar Chemistry
Physical and Chemical Properties
CH3CHOHCH3.
60.10.
0.877 g/mL @ 20°C.
2.1.
-88°C.
83°C.
33 mm Hg.
11.7°C (53°F) CC.
399°C (750°F).
Section 10:
Appearance
Odor
Odor Threshold
Solubility
Evaporation rate
Partition Coefficient
pH
LEL
UEL
Clear, colorless liquid.
Alcohol odor.
N/A
Completely soluble in water.
> 1 (Butyl acetate = 1).
N/A. (log POW).
N/A.
2%.
12.7 %.
N/A = Not available or applicable
Stability and Reactivity
Stability: Stable under normal conditions of use. Avoid heat and ignition sources.
Incompatibility: Oxidizing materials, caustics, aluminum, metal, oleum, chlorinated compounds.
Shelf life: Fair shelf life, store in a cool, dry environment.
Section 11:
Toxicology Information
Acute Symptoms/Signs of exposure: Eyes: Stinging pain, watering of eyes, inflammation of eyelids and conjunctivitis. Skin:
Insensitivity to pain, feel of coolness or cold, skin looks white and feels hard and cold. Ingestion: Breath has sweet, organic
odor, mental confusion, drowsiness, nausea, vomiting and headache. Inhalation: Rapid irregular breathing, headache, fatigue,
mental confusion, nausea and vomiting, giddiness and poor judgment, convulsions and death.
Chronic Effects: Repeated/prolonged skin contact may cause dryness or rashes.
Sensitization: none expected
Isopropyl Alcohol: LD50 [oral, rat]; 5045 mg/kg; LC50 [rat]; 16,000 mg/l (4hours); LD50 Dermal [rabbit]; 500mg/24H Mild
Material has not been found to be a carcinogen nor produce genetic, reproductive, or developmental effects.
Section 12:
Ecological Information
Ecotoxicity (aquatic and terrestrial):
Toxic to aquatic and terrestrial plants and animals. Do not release into environment.
Section 13:
Disposal Considerations
Check with all applicable local, regional, and national laws and regulations. Local regulations may be more stringent than
regional or national regulations. Small amounts of this material may be suitable for sanitary sewer or trash disposal.
Section 14:
DOT Shipping Name:
DOT Hazard Class:
Identification Number:
Transport Information
Isopropano.l
3, pg II.
UN1219.
Section 15:
EINECS: Listed (200-661-7).
TSCA: All components are listed or are exempt.
Canada TDG:
Hazard Class:
UN Number:
Isopropanol .
3, pg II .
UN1219.
Regulatory Information
WHMIS Canada:B2, D2B: Flammable liquid, Toxic material: eye irritant.
California Proposition 65: Not listed.
The product has been classified in accordance with the hazard criteria of the Controlled Products Regulations and the MSDS
contains all the information required by the Controlled Products Regulations.
Section 16:
Other Information
Current Issue Date: December 21, 2011
Disclaimer: Scholar Chemistry and Columbus Chemical Industries, Inc., (“S&C”) believes that the information herein is factual but is not intended to be all
inclusive. The information relates only to the specific material designated and does not relate to its use in combination with other materials or its use as to any
particular process. Because safety standards and regulations are subject to change and because S&C has no continuing control over the material, those
handling, storing or using the material should satisfy themselves that they have current information regarding the particular way the material is handled, stored
or used and that the same is done in accordance with federal, state and local law. S&C makes no warranty, expressed or implied, including (without
limitation) warranties with respect to the completeness or continuing accuracy of the information contained herein or with respect to fitness for any
particular use.
© 2008, Scholar Chemistry. All Rights Reserved.
©2012, Ward’s Natural Science
All Rights Reserved, Printed in the U.S.A.
12/21/2011
US: www.wardsci.com
Canada: www.wardsci.ca
250-7456 v.5/12
Page 37