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LABORATORY MANUAL
SBU 3023
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
SBU3023 BIOLOGY II
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
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
Principle of Mendel
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.
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.
SBU3023 BIOLOGY II
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
Experimental procedures:
SBU3023 BIOLOGY II
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.
2
SBU3023 BIOLOGY II
DNA sequence
Codon
mRNA
in
Amino Acid
CGT
CCA
CGT
CCA
SBU3023 BIOLOGY II
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
2a
2b
Filament is unbranched 3
Filament is branched Chaetophora (Cholophyceae)
3a
3b
4a
4b
5a
5b
6a
6b
Organism is a unicell 7
Organism is a colony 13
7a
7b
8a
8b
9a
9b
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
SBU3023 BIOLOGY II
13a
13b
14a
14b
15a
15b
16a
16b