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Faculty of Science and Mathematics

LABORATORY MANUAL

SBU 3023
BIOLOGY II

LABORATORY GUIDELINES AND PROCEDURES


SBU3023 BIOLOGY II

ATTENDANCE
1. Attendance at all practical classes is compulsory. Students who fail to
attend any of the practical session have to provide a written notification
or medical certification.
2. All students who are attending a practical session must sign the
attendance sheet, else the student will be considered absent.

PREPARATION
1. Students need to be well prepared and planned their experimental
exercise thoroughly before the practical session.

PRACTICAL REPORT
Students must write a practical report for each of the practical exercise. All of
the report must be hand in by the end of the week of the practical session to the
lab.
a) Diagram (a practical exercise that involves observing slides through
microscopes)
b) A practical report must consist of:
i)
Title
ii)
Practical exercise
iii) Objective
iv) Prediction/ Hypothesis
v)
Results/Findings/Observations
(includes any table, graph, description, or diagram)
vi) Discussion

Discuss and analyse the results obtained from the practical


exercise. Provide a justification of your finding in the practical
exercise.
vii) Conclusion
viii) References
All of the references and resources used in the report need to be
cited properly.

SBU3023 BIOLOGY II

CELL DIVISION MITOSIS & MEIOSIS


Laboratory exercise 1:
Objective:

Investigate the cell divisions in mitosis and meiosis

To understand cell cycle stages via mitosis and meiosis

Introduction:
Organism produces its offspring through the process of reproduction, which involved
cells division. Mitosis and meiosis are both cell division mechanisms. However, the
outcomes of each mechanism are different. You will study and review the cell division
mechanisms at different mitosis and meiosis stages using the Allium cepa root tips and
some prepared slides.
Material and Apparatus:
A. Mitosis
1. Allium cepa root tips ( treated and untreated root tips)
2. Microscope
3. Glass plate
4. Glass slide
5. Pin
6. Cover slip
7. Filter paper
8. Alcohol lamp
9. Acidic Aseto Orsein

Note: The root is treated with paradicholorobenzene in 3 hours

Root tips are preserved by using pure ethyl alcohol and glacial acetic acid (3:1)
C.

B. Meiosis
1. Microscope
2. Prepared slides

SBU3023 BIOLOGY II

Methods:
1. Place 3 or 4 root tips in several drops of acidic Aseto-orsein (9 acidic Aseto
Orsein: 1 10% HCl) on the glass plate.
2. Heat the glass plate by using alcohol lamp for 2-3 minutes. (Careful not let the
glass plate to burn over).
3. Cover with the glass plate and leave it for 10 minutes.
4. Place a root tip on a clean glass slide. Cut into 1 to 2 mm from the end of the tip
and remove the remaining part.
5. Squash the root tip to the tiny pieces by using a pin.
6. Add 1 or 2 drops of Aseto-orsein onto the slide. Carefully wipe any remaining
Aseto-orsein around the specimen with filter paper.
7. Cover the glass slide with a cover slip (do not allow it to dry), then gently knock
the cover slip by using a short piece of wood (matches). This step is performed to
separate the root cells.
8. Slowly heat the glass slide by placing it on the alcohol lamp for a while. (Careful
not let the glass slide to burn over). Then, place the glass slide between filter
papers and press the cover slip gently for a complete cells separation.
9. Examine the slides under the microscope.

Note: Apply step 4 to step 9 on treated and untreated root tips. Apply step 9 on the
prepared slides of the meiosis stages.

Observation:

Observe the slides and identify the cell division stages that you can see. Describe and
draw each of the observed stages in your report.

SBU3023 BIOLOGY II

MENDELS INHERITANCE LAW


Laboratory exercise 2:
Objective:

Principle of Mendel

To understand the concept of the Mendelians Inheritance Laws

Introduction:
Gregor Mendel was the first person who studied the genetic materials transmission in
inheritance. He found the inheritance principles that were the basic of modern genetics.
These inheritance principles were also known as Mendels First Law: Segregation, the
Second Law: Independent assortment.
Mendels Law of Segregation:
During the gametes formation, the two alleles of each trait separate (segregate), and then
unite at random, one from each parent, at fertilisation.
Mendels Law of Independent assortment:
During gametes formation, different pairs of alleles segregate independently from each
other.

Material and apparatus:


1. Maize kernels
a. Ratio 3:1
b. Ratio 1:1
c. Ratio 9.3.3.1
d. Ratio 1:1:1:1
Methods:
A. Principle of segregation
1. You will be given maize kernel, which resulted in a cross of the contrasting traits.
Observe the characteristics appeared on the maize kernels (a) and (b). Observe the
characteristics appeared on the maize kernels (a) and (b). Maize kernel (a) is
obtained from F1 cross which was cross between purple seed and yellow seed.
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SBU3023 BIOLOGY II

While, maize kernel (b) is obtained from a test cross of F1 generation (purple X
yellow).
2. Determine the dominant phenotype
3. Count the number of every phenotypes
4.

Count the expected number of every phenotype

B. Principle of independent assortment


1. You will be given the maize kernels, which resulted from a cross of two pairs of
contrasting traits. Observe the characteristics appeared on the maize kernels (c)
and (d). Maize in kernel (c) is obtained from the F1 cross which was a cross
between purple and round seed with yellow and wrinkled seed. While maize
kernel (d) is obtained from a test cross of F1 generation, (purple and round seed X
yellow and wrinkled seed).
2. Determine the dominant and recessive phenotype
3. Count the numbers of every phenotype
4. Count the expected numbers of every phenotype
Observation:
Record your observation in a table form. Review and write a report about your
observation and understanding on the Mendelians Laws.

SBU3023 BIOLOGY II

From DNA to Protein


Laboratory exercise 3:
Objectives:

DNA, mRNA and protein.

To build and appreciate the DNA structure.


To apply and determine the product of transcription and translation processes.

Introduction:
Deoxyribonucleic acid (DNA) holds heredity information. A DNA molecule consists of
two long chains of nucleotides that coiled into a double helix. The chains composed of
nucleotides, which each has phosphate groups, a deoxyribose sugar and a nitrogenous
base. These two chains held together by the hydrogen bonds.
The DNA puzzle kit includes the following and should be carried out in the following
order:
1. DNA: the genetic code
2. The Code Transcribed and Translated

A. DNA: The genetic code

Materials and apparatus:


24 deoxyribose units (red)
24 phosphate units
4 adenine units
8 cytosine units
8 guanine units
4 thymine units

Experimental procedures:

I. Constructing DNA nucleotide


1. Attach one phosphate unit to the free bond from CH2 deoxyribose molecule.
2. At the right hand side of the oxygen atom, there is a carbon atom with one free
bond. Attach the adenine to the bond. The structure you have constructed is
called adenine nucleotide.
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SBU3023 BIOLOGY II

3. Construct nucleotide having guanine, cytosine and thymine.

II. Joining nucleotide through covalent building


1. Select an adenine nucleotide and a guanine nucleotide that had been prepared
earlier.
2. Attach both nucleotides together at phosphate unit from one nucleotide to 3
end of another nucleotide.
3. Repeat step 2 until you get a DNA chain consisting of six nucleotides.

III. Base pairing of DNA molecules


1. Separate the DNA chains constructed approximately 30cm apart.
2. Built two new DNA chains on each separated stand, which complement and
anti-parallel with each other.

B. The code transcribed and translated

Materials:
12 deoxyribose units (red)
12 ribose units (pink)
24 phosphate units
4 adenine units
8 cytosine units
8 guanine units
2 thymine units
2 uracil units

Experimental procedures:

1. Construct a DNA chain having the following sequences: CGT CCA CGT CCA
2. Construct a RNA chain with is complement and antiparallel with the DNA chain
built in step 1. Remember, in RNA base thymine is replaced by base uracil.
3. By referring to codon table for mRNA, fill in the table below with their
respective translated mRNA sequences and transcribed amino acids.
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SBU3023 BIOLOGY II

DNA sequence

Codon
mRNA

in

Amino Acid

CGT
CCA
CGT
CCA

1. What are the differences between DNA and RNA?


2. Which direction does DNA replication occurs?
3. What are the differences between a leading and lagging strand in DNA
replication?
4, State the base sequences of start codon and stop codon.

SBU3023 BIOLOGY II

TAXONOMY: ORGANISMS CLASSIFICATION


Laboratory exercise 4:
Objectives:

Classify invertebrate into the taxa

to determine organisms using the dichotomous key.

Introduction:
Dichotomous key is one of a common method used to classify an unknown organism. The
observation on the characteristics, such as structure, behaviour, of the unknown organism
helps to recognise the organism. However, careful observations are required to induce the
right name of the organism. Dichotomous means divided into two parts. Therefore,
dichotomous keys always offer two choices for each steps. Each key describes a
characteristic of a particular organism or group of organisms.

Materials
1. Prepared slides
2. Microscope

Procedures:
1. Examine the prepare slides using the microscope. Examine your material first using
the lower power objective (i.e. 10X); then use a higher power objective (i.e. 20X or
40X). Because the objectives are parfocal, you need to use only the fine focus knob to
fine tune your image. Never use the coarse adjustment to focus downward. Replace
and remove a slide only after the lowest power objective has been rotated into
viewing position.
2. Use the dichotomous key shown in the Appendix 1 (Darley M., 2003) to recognise the
organisms.
3. Record the name of the organisms and draw the features of each organism from your
observation. Note the power objective of the microscope used in the observations.

SBU3023 BIOLOGY II

Appendix 1
A Dichotomous Key of Pond Life
1a
1b

Organism is a filament (a liner series of cells) 2


Organism is a unicell or a colony (flat or spherical) 6

2a
2b

Filament is unbranched 3
Filament is branched Chaetophora (Cholophyceae)

3a
3b

Choloplast fills cell: filaments often end in H piece Microspora (Cholophyceae)


Chloroplast has a distinctive shape 4

4a
4b

Chloroplast is in the form of a spiral Spirogyra (Charophyceae)


Chloroplast is not in the form of a spiral 5

5a
5b

Two star-shaped chloroplast in each cell Zygnema (Charophyceae)


Two thin, plate-like chloroplasts in each cell Mougeotia (Charophyceae)

6a
6b

Organism is a unicell 7
Organism is a colony 13

7a
7b

Organism cell does not contain flagella 8


Organism cell is motile with flagella 11

8a
8b

Organism cell is green 9


Organism cell is golden brown and elongated (pinnate diatom) Nitzschia
(Strminopila)

9a
9b

Cell is round, not divided into halves Chlorococcum (Chlorophyceae)


Cell is not round, often appears to be divided into halves with nucleus in the middle of
the cell, dark green (desmids, Charophyceae) 10

10a

Cell is elongated without a constriction in the middle; cell may be slightly curved
(like a banana) Closterium
Cell has an obvious constriction in the middle; cell highly ornate with several lobes
and secondary lobes Micrasterias

10b

11a
11b

Cell is green 12
Cell is not green, with two sub-apically inserted flagella; may be blue-green, brown,
radish brown cryptomonads (genus unknown)

12a
12b

Cell elongated with one long flagellum Euglena (euglenid, Euglenozoa)


Cell is oval with two flagella Chlamydomonas (Chlorophyceae)
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SBU3023 BIOLOGY II

13a
13b

Colony is spherical and motile 14


Colony is non-motile 15

14a
14b

Colony is green Volvox (Chlorophyceae)


Colony is yello-brown Synura (Chrysophyta)

15a
15b

Colony is spherical Coelastrum (Chlorophyceae)


Colony is not spherical 16

16a
16b

Colony is a round, flat plate Pediastrum (Chlorophyceae)


Colony has 2 or 4 cells with spines on the corners Scenedesmus (Chlorophyceae)

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