CAPE UNIT 1

BIOLOGY
PRACTICAL HANDBOOK

POLYTECHNIC SIXTH FORM

GOVERNMENT

Ms TESLA GOULD

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Contents
SECTION 1 ..................................................................................................................................................... 4
1. WRITING LABORATORY REPORTS ..................................................................................................... 4
OBSERVATION TYPE LAB REPORTS ....................................................................................................... 4
EXPERIMENT TYPE LAB REPORTS .......................................................................................................... 5
PLAN ANDDESIGN LAB REPORTS........................................................................................................... 6
2. MICROSCOPY..................................................................................................................................... 7
TIPS........................................................................................................................................................ 8
4. CALCULATING THE MAGNIFICATION .............................................................................................. 10
5. BIOLOGICAL DRAWINGS ................................................................................................................. 11
6. MAKING A WET MOUNT ................................................................................................................. 11
SECTION 2 – MARKING GUIDELINES ........................................................................................................... 12
1. DRAWING SKILL ............................................................................................................................... 12
2. MEASUREMENT AND MANIPULATION SKILL .................................................................................. 13
3. ANALYSIS AND INTERPRETATION SKILL........................................................................................... 14
4. OBSERVATION/RECORDING/REPORTING SKILL .............................................................................. 15
5. PLANNING AND DESIGN SKILL......................................................................................................... 16
6. THE INVESTIGATIVE PROJECT.......................................................................................................... 17
SECTION 3 – PRACTICALS ............................................................................................................................ 19
1. Wet Mount of Onion and Rhoeo Epidermal Cells and Epidermal Peels ......................................... 19
2. Comparing Solubility of Substances in Liquids of Different Polarities ............................................ 20
3. Food Tests ....................................................................................................................................... 22
Or Alternatively: June 2009, Paper 3/2 Question 1 ................................................................................ 24
4. Quantitative Investigation and Comparison of Reducing Sugars and Starch ................................. 27
5. Diagrams and Electron Micrographs of Cell Membranes and Organelles in Animal and Plants .... 27
6. Electron Micrographs and Prepared slides of Prokaryotic Cells ..................................................... 27
7. Tissue Plan and Detailed Diagrams of Tissues in Dicotyledonous Plant Roots and Stems ............. 28
8. Investigating the Effect of Immersion into Solutions of Differing Water Potentials on Plant Cells 29
9. Effect of Temperature on Enzyme Reactions ................................................................................. 30
10. Effect of Substrate Concentration on Enzyme Reactions ........................................................... 31
11. Stages of the Cell Cycle from a Root Tip Squash and Prepared Slides ........................................ 32
12. Meiosis Model............................................................................................................................. 32

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13. Stages of Meiosis from Prepared Slides of the Testis or Anther ................................................ 33
14. Drawing of the Anther and a Plant Embryo Sac.......................................................................... 33
15. Seed Samples and Mendelian Genetics ...................................................................................... 34
16. Drawing of a Mammalian Testis and Ovary ................................................................................ 37
17. Planning and Designs .................................................................................................................. 37
18. The Investigative Project............................................................................................................. 37

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SECTION 1

1. WRITING LABORATORY REPORTS

Headings for ALL laboratory reports should appear in capital letters and must be underlined with a single
line. ALL diagrams must be drawn in pencil with the title for each diagram appearing at the bottom in
pencil, capital letters and underlined with a single line.

OBSERVATION TYPE LAB REPORTS

Observation type practicals are drawing labs. All sub-headings must be written as shown in capital
letters and underlined. The report should take the following format:

DATE:

LAB NO.:

TITLE:

SYLLABUS REFERENCE:

INTRODUCTION:

Introductions should provide some background information on which the practical activity is based. It
should include definitions where possible.

AIM:

APPARATUS AND MATERIALS:

METHOD:

Must be written in the past tense.

OBSERVATIONS:

SUMMARY:

A summary demonstrates your understanding of the specimen viewed. Your observations should relate
to and compare your expectations based on theory/notes and textbook information to what was
actually seen. It is not meant to be a repetition of your introduction.

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EXPERIMENT TYPE LAB REPORTS
Experiment type practicals are those in which scientific experiments are done to investigate different
and known phenomena. The report should take the following format:

DATE:

LAB NO.:

TITLE:

SYLLABUS REFERENCE:

INTRODUCTION:

Introductions should provide some background information on which the practical activity is based. It
should include definitions where possible.

AIM:

APPARATUS AND MATERIALS:

METHOD:

Must be written in the past tense.

RESULTS:

Should accurately represent ALL the data you collected while conducting the experiment. The data must
be presented in a suitable format such as a table with an appropriate title in capitals and underlined
and, appropriate column headings with units where needed. Some experiments produce data that can
be pictorially illustrated with a graph. A graph must follow ALL the usual mathematical rules to be
considered complete. A graph must have a title appearing at the top in capital letters, labelled axes with
units, a scale, a key where necessary and should maximise the graph page.

DISCUSSION:

Definitions may be re-stated. The discussion requires a thorough explanation of your findings including
all trends observed in your data as presented in your tables and graphs. The chemistry behind every
chemical analysis/reaction must be thoroughly explained including the reasons for colour changes,
movement, gases evolved, etc. Each intermediate or product must be identified by name, type of
reaction occurring and possible enzymes involved, etc.

Your main findings should be re-stated in the final paragraph of your discussion.

CONCLUSION:

Should answer the experiment AIM you stated. Anything else is NOT a conclusion. It is not a paragraph
and does not require explanations or definitions.

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PLAN ANDDESIGN LAB REPORTS

Please ensure that your lab report follows the following format:

PROBLEM STATEMENT:

HYPOTHESIS:

AIM :

APPARATUS AND MATERIALS:

PROCEDURE :

 Controlled variable must be stated

 Manipulated variable must be stated
 Number of repetitions stated
 Precautions stated
 Method precise and easy to follow

EXPECTED RESULTS :

 Expected changes example colour change that shows a positive result

 Expected results stated

TREATMENT OF RESULTS :

 Stated how the results will we treated and used, for example: graph, calculations, etc
 All Possible limitations of this experiment stated

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 2. MICROSCOPY
Source: https://www.pinterest.com/pin/531635930982961626/

Glossary source: http://www.microbehunter.com/parts-of-a-compound-microscope/

The following list of terms can also be found in the glossary:

 Condenser: This is a system of different lens elements which is mounted beneath the
stage of the microscope. It contains an iris diaphragm which controls the diameter of
the light beam. The light beam should be adjusted to be larger or equal to the numerical
aperture of the objective in use. Condensers can be moved up and down. The normal
operating position is up. The purpose of the condenser is to ensure that the objective is
able to provide the maximum resolution. By adjusting the aperture diaphragm, it is also
possible to Control depth of field and contrast. While this also changes light intensity,
this should only be considered a side-effect (should not be used to control light)
 Base: This is the bottom part of the microscope, it contains the lamp.
 Coarse Focus: Also referred to as rough focus, this knob raises and lowers the
microscope stage quickly. It should only be used in connection with the low
magnification lenses.
 Eyepiece Lens: Also known as ocular lenses, they magnify the image of the objective.
The eyepiece is the lens into which a person looks into when observing. The total
magnification of a microscope is calculated by multiplying the magnification of the

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 objective by the magnification of the eyepiece. Many eyepiece lenses have a
magnification of 10x ot 15x.
 Fine Focus: This focus knob moves the stage up and down in small steps. It is used to
focus at different layers of the specimens.
 Head: This is the top part of the microscope. It carries the eyepiece(s) and other optical
elements. There are several different types of heads: a monocular head is designed to
carry only one eyepiece, a binocular head carries two (but does not give stereoscopic
vision in compound microscopes) and a trinocular head is designed to carry a camera as
well.
 Mechanical Stage: This type of stage is equipped with a slide holder and two knobs to
turn. One knob moves the stage backwards and forwards, the other one moves the slide
sideways.
 Nosepiece (or revolving nosepiece, turret): This part carries the low, medium and high
power objective lens. It can be rotated.
 Objective Lens: This is a highly magnifying lens system, it is located close to the
specimen to be observed. The image of the objective is then magnified again by the
ocular lens which is close to the eye.
 Stage: This is the flat surface on which the slides are placed on. It can be moved up and
down for focusing.
 Stage Clips: These are clips that hold the slide.

TIPS
1. Always remove the microscope from its case by properly supporting it with one hand on the arm
and one hand on the base of the microscope
2. Place the slide on the microscope stage with the specimen directly over the lens from which the
light is shining through. Makes it easier to find the specimen as the coarse focus is adjusted
3. Always start by finding the specimen with the low power lens. Once found, centre the specimen
and switch to a higher power lens
4. Keep both eyes open at the same time to reduce eye strain
5. Reducing the light often makes it easier to see the specimen as too much light makes the
specimen appear washed out

- Place prepared slide on the stage with specimen on centre of stage opening
- Set diaphragm at the maximum opening
- Turn nosepiece with low objective in direct line with tube (Clicks into position)
- Turn on sub-stage light
- Turn coarse adjustment downward until the objective is very close to the slide while
looking at microscope from side
- Make sure objective does not touch the slide
- Adjust the diaphragm to produce the sharpest image

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 - Use fine adjustment to obtain sharpest image
For High Power

- Begin observation of specimen under low power

- Turn nosepiece until high power objective clicks into position
- Use fine adjustment only to bring image into sharp focus
- Determine diameter of specimen in terms of eyepiece graticule units
- Calculate actual size of specimen using calibration with stage micrometer ( calibration
would have been done during the term) or using diameter of field of view method

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 4. CALCULATING THE MAGNIFICATION

CALCULATING MAGNIFICATION FROM LIVE SPECIMEN

Magnification = Size of Drawing

Size of specimen

CALCULATING MAGNIFICATION FOR DIAGRAMS FROM MICROSCOPE

A stage micrometre ruler is 1 cm in length with 100 divisions.

Each division on the graticule ruler inside your lens is called an ocular division (o.d.)

The smallest distance of the stage micrometre = 1 mm

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1 mm = 1000 μm

Microscope Magnification = ocular magnification x objective magnification

Ocular Lens Objective Lens Microscope Magnification

X 10 Low power ( X 4) X 40
X 10 Medium power ( X 10) X 100
X 10 High power ( X 40) X 400

Microscope Magnification Value of Each Graticule Division

X 40
X 100
X 400

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 5. BIOLOGICAL DRAWINGS

TIPS:

1. Diagrams, labels, calculations for magnification and titles must be written in pencil
2. NO SHADING, COLOURING, SKETCHING or cross hatching
3. Diagram must be a reasonable size. Diagram must occupy the upper left hand section of your
page and must be as large as possible leaving sufficient space for labels to be written to the right
of it
4. Titles must be written in capital letters and satisfy all the criteria stated in the mark scheme
provided; uppercase and underlined including view, specimen name and magnification
5. Label lines must be straight, horizontal and parallel with no arrowheads or dots and be fully
justified
6. Line touching structure being labelled
7. The lines on your diagram must be clean, continuous and of even thickness
8. No excessive details on plan drawing diagrams
9. The details of your diagram must represent as accurately as possible the proportions of the
specimen being viewed

6. MAKING A WET MOUNT

1. With a medium dropper/teat pipette, place a drop of water in the centre of a clean slide
2. Using forceps, peel off thin epidermis that lines the inside (concave side) of the onion
3. Cut a small piece (3 to 5 mm wide and about 10mm long) of epidermis with sharp
scalpel/scissors
4. Place the piece of the specimen on the drop of water on the slide
5. Place one edge of a clean cover slip on the edge of water but not touching the specimen
6. Support the opposite edge of cover slip with the mounting needle
7. Slowly lower cover slip until it lies flat
8. Place a drop of stain at one edge of the cover slip
9. Draw the stain under the cover slip by touching a piece of paper towel to opposite edge
10. If any air bubbles present, push them out gently by pressing on the cover slip with back of the
mounting needle
11. Mop up any solution on top of cover slip with tissue
12. Periodically replenish solution by adding with pipette to edge of cover slip

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 SECTION 2 – MARKING GUIDELINES

These are samples of mark schemes that I actually use. Please refer to them as often as needed.

1. DRAWING SKILL

Criteria Details Maximum Student

Mark Mark

Clarity Clean, continuous lines 4-5=3

Lines of even thickness 3 =2
No excessive details on plan drawing 2-1=1
No shading or cross hatching
Drawing is a reasonable size

Label lines Horizontal and parallel with no arrowheads or dots 1

Line touching structure being labelled 1
Labelled in lowercase script in pencil 1

Labels and At least 6 labels correct with annotations as necessary 6-8=3

Annotations Labels incorrect but annotations correct (-1 penalty) 3–5=2
where Labels correct but no annotations (-1 penalty) 1-2 = 1
necessary

Title Uppercase and underlined including view, specimen name and 1

magnification

Accuracy Diagram resembles the specimen 3–5=2

Correct shape of specimen 1-2=1
Correct proportions of tissues
Parts positioned correctly
Any cut edges represented by a double line

TOTAL 12

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 2. MEASUREMENT AND MANIPULATION SKILL

Example Skill: Use of the Microscope

CRITERIA Maximum Student

Mark Mark
The microscope was removed from its case with one hand on its arm 1
and the other on its base
Slide was placed on the stage and stage clips positioned to hold it firmly 1

The slide was handled with care 1

Both eyes were kept open while looking through the binocular lens 1

Specimen was found at low power by correctly adjusting the coarse 1

focus and stage
Specimen was correctly brought into focus by adjusting the fine focus 1

The ocular lens was turned until the eyepiece micrometer was 1
positioned along the widest area of the specimen
Stage was adjusted correctly until the zero line of the eyepiece micro- 1
meter was directly in line with the edge of the widest area of specimen
Number of ocular divisions on the eyepiece micrometer representing 1
the length of the widest area of the specimen was accurately counted
Student accurately calculated the actual size of the specimen in μm at 1
low power by multiplying by 25
Student accurately measured the widest area of plan drawing in cm to 1 1
decimal place and converted this figure to μm
Magnification of drawing was accurately calculated to 1 decimal place 1

TOTAL 12

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 3. ANALYSIS AND INTERPRETATION SKILL

Criteria Details Max Student

Mark Mark
Analysis Results
*Complete set of results from quantities mentioned in method. 2

Interpretation Discussion
*Complete set of calculations. 2
*Statement of observations and trends.

*Interpretation of calculated values and trends related to the 2

data presented in the results.

Limitations / Sources of Errors

At least one accurate limitation stated. 1
At least one accurate source of error stated 1

Reflection
*Explains relevance of the experiment to real life. 1
*States the impact of knowledge gained from the experiment. 1
*States how the experiment may be improved or changed to get 1
better or more accurate results.

Conclusion
Clearly stated and related to the aim of the Plan and Design. 1

TOTAL 12

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 4. OBSERVATION/RECORDING/REPORTING SKILL

Criteria Details Maximum Student

Mark Mark
Format *Neat, correct sequence (title, syllabus objective, introduction, 1
aim, apparatus/materials, results, discussion and conclusion)
*Sub-headings clearly stated in capital letters and underlined 1
*All apparatus and materials included 1
*Method
-Correct sequence of tasks written in past tense 1
-includes precautions, a control and a replication 1

Language Concise, clear, written in past tense and in standard English 1

Results Data organised and presented in a Table

*Correct number of rows and columns with appropriate headings 1
and units as necessary
*Appropriate title above table in capitals and underlined 1

Graph used as a pictorial representation of data

Format of graph
*Appropriate title above graph and underlined 4-5 = 2
*All axes labelled with units included 2-3 = 1
*Manipulated/ Independent variable on x-axis ≤1=0
*Appropriate scale
*Appropriate key

Construction
*All points accurately plotted in pencil 1
*Neat, best fit curves drawn 1

TOTAL 12

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 5. PLANNING AND DESIGN SKILL

Criteria Details Maximum Student

Mark Mark

Hypothesis Specific 1
Measurable – testable as a student exercise 1

Aim Related to the hypothesis, specific and relevant 1

Apparatus Applicable to method 1

and
Materials

Procedure Control – stated or implied 1

Specific amounts or quantities and/or timeframe stated 1
Logical sequence of steps 1
Instructional point form in present tense 1
At least 1 repetition 1

Results Expected results stated 1

Interpretation of results stated 1

Source of At least 1 stated 1

errors/
Limitations

TOTAL 12

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 6. THE INVESTIGATIVE PROJECT

Investigative Project

Plan and Design Format – P&D IA

Please ensure that your P&D report follows the following format: Marks stated in [ ] brackets

PROBLEM STATEMENT

HYPOTHESIS [1]

AIM [1]

APPARATUS AND MATERIALS [1]

PROCEDURE [5]

 Controlled variable [1]

 Manipulated variable [1]
 Number of repetitions stated [1]
 Precautions stated [1]
 Method precise and easy to follow [2]

EXPECTED RESULTS [2]

 Expected changes example colour change that shows a positive result [1]
 Expected results stated [1]

TREATMENT OF RESULTS [2]

 Stated how the results will we treated and used, for example: graph, calculations, etc [1]
 All Possible limitations of this experiment stated [1]

********** TOTAL = 12 MARKS ***********

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Investigative Project

Report Format – Analysis and Interpretation IA

Please ensure that your report follows the following format: Marks stated in [ ] brackets

TITLE: Social and Preventative Medicine – Diabetes and Obesity

SYLLABUS REFERENCE: Unit 2 Module 3 Topic 3 Objective 3.1

INTRODUCTION

AIM

APPARATUS AND MATERIALS

METHOD

RESULTS [2]

DISCUSSION

 Stated and Explained trends [2]

 Accurate interpretation of results and trends identified [2]
 Limitations identified [1]
 Sources of error identified [1]

REFLECTIONS

 Relevance to real life stated [1]

 How the knowledge gained in this experiment impacted you [1]
 How do you think the experiment can be changed or improved [1]

CONCLUSION [1]

********** TOTAL = 12 MARKS **********

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SECTION 3 – PRACTICALS

1. Wet Mount of Onion and Rhoeo Epidermal Cells and Epidermal Peels

AIM: To prepare wet mounts of onion and Rhoeo epidermal cells and make diagrams

APPARATUS AND MATERIALS:

Slides, coverslips, mounting needle, methylene blue, distilled water, razor blade, forceps, tissue,
onion and Rhoeo leaf.

METHOD:

With a medium dropper/teat pipette, place a drop of water or stain in the centre of a clean
slide. Make a small cut in the epidermis of the specimen with the razor blade. Using the
forceps, peel off the thin epidermis from the cut surface of the specimen. For the onion, a small
piece (3 to 5 mm wide) is cut with the razor blade from the concave side before using the
forceps to peel it off. Place the piece of epidermal tissue on the drop of water on the slide.
Ensure that there is no folding or overlapping areas of tissue. Place a drop of the stain on the
epidermal tissue and allow a minute or two for absorption. Place one edge of a clean cover slip
against the edge of the drop of liquid on the slide (not on the specimen). Support the opposite
edge of cover slip with the mounting needle and slowly lower the cover slip until it lies flat over
the specimen. Touch a piece of tissue to the outer edge of the coverslip to absorb and remove
any excess liquid on the slide. If any air bubbles are present, push them out gently by pressing
on the cover slip with back of needle and giving them a light tap (don’t break the coverslip
please). Place slide on the stage of the microscope and make a detailed diagram of 2-4 cells
under high power.

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 2. Comparing Solubility of Substances in Liquids of Different Polarities

AIM: To compare the solubility of substances in liquids of different polarities

APPARATUS AND MATERIALS:

9 test tubes, 9 rubber stoppers, test tube rack, 3 mL glycerol, iodine, sodium chloride, 15 mL
distilled water, 15 mL ethanol, 15 mL hexane

METHOD:

Prepare a data table that will allow you to record your observations for each solute-solvent
combination. (see below)

Data Table:

Solvent \ Solute Glycerol Iodine Sodium chloride

Distilled Water

Ethanol

Hexane

Place 5 ml of water in each of three test tubes. Add 1 ml of glycerol to the first test tube, two
small iodine crystals to the second and a few small sodium crystals to the third. (CAUTION: Do
not touch the iodine crystals!) Stopper all three test tubes, shake them vigorously, and allow
them to stand for 1 minute. Observe the contents of each test tube and record your
observations. Repeat Step 2 using ethanol instead of water. (Is ethanol polar or non-polar?).
Repeat Step 2 using dichloromethane instead of water. (Is dichloromethane polar or non-
polar?). Record if the solute is insoluble, slightly soluble or very soluble in the particular
solvent. (Remember to use miscible and immiscible to describe the solubility of two liquids).

Data Analysis:

1. In which solvent is glycerol most soluble? In which solvent is it the least soluble?

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2. In which solvent is sodium chloride most soluble? In which solvent is it the least soluble?
3. In general, in what type of solvent (non-polar, moderately polar, or highly polar) are polar
solutes most soluble? Explain why.
4. In which solvent is iodine the most soluble? In which solvent is it the least soluble?
5. When interactions are weak, then the molecules can be easily separated from each other.
Predict whether a non-polar compound or a polar compound is more easily separated into
isolated molecules. Explain.

**Note: all the interactions are due to + and - charges attracting each other.

6. What is the nature of the attracting charges in ionic compounds?

7. What is the nature of the attracting charges in polar compounds?
8. What is the nature of the attracting charges in non-polar compounds?
9. What is meant by an induced dipole?
9. Draw 2 Br2 molecules, and show a moment in time when they have induced dipoles that
cause them to be attracted to each other.
10. Polar compound interactions are termed dipole-dipole because each already is a polar
substance. Draw 2 HCl molecules and show how they would be arranged when interacting.
11. Water, and some other compounds, have strong dipole-dipole interactions called hydrogen
bonds. Draw 2 H2O molecules with a hydrogen bond between them.
12. Ions interact strongly with polar compounds. That is why salts, which are made of ions, can
dissolve in water. Draw a picture that shows how this occurs.
13. Which part of a water molecule is attracted to a cation (positive ion)?
14. Why won't non-polar compounds dissolve in water too? Or why doesn't salt (NaCl) dissolve
in hexane?
15. Explain why NaCl won't dissolve in hexane (a non-polar solvent) using a similar thought
process.
16. Within each pair of compounds, pick the one more likely to be soluble (mix) with water.
Explain.
a) NH3 (polar) and CH4 (non-polar)
b) HCl (polar) and ICl5 (non-polar)
17. You observe that a substance dissolves in hexane but not in water. What kind of substance
do you think it is, polar or non-polar? Explain.
18. Salt and sand are mixed together as solids. Describe what happens at the molecular level,
when you add water.
19. Is water polar? Explain.

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3. Food Tests
AIM: To determine the biomolecules present in different food samples

APPARATUS and MATERIALS:

Droppers, NaOH, HCl, copper sulphate, Benedict’s reagent, iodine/KI solution, 1% starch, 1%
egg albumin, oil, ethanol, 1% sucrose solution, 1% glucose solution, chicken stock, small beaker,
syringe, 10 ml measuring cylinder, labels, distilled water, test tubes, test tube rack, test tube
holder, Bunsen burner, tripod stand, wire gauze, large beaker.

METHOD:

Using the procedure outlined for the various food tests, test the food samples identified in the
table for the biomolecule stated. Record all colour changes and observations as accurately as
possible. Record all results in a suitable tabular format.

TEST FOR STARCH

Place a small sample of the food item to be tested in a test tube. Add 2 drops of the iodine in
potassium iodide solution and swirl to mix. Record observations.

TEST FOR LIPIDS

Using the measuring cylinder, measure approximately 1ml of ethanol and pour into a test tube.
Measure 2 ml of distilled water using the measuring cylinder. Pour a small amount of oil into it.
Cover the mouth of the test tube with your thumb and shake vigorously. All the 2ml of water to
it, shake and record all observations. Allow to stand for a few minutes and record observations
again.

PROTEIN TEST
Measure 1ml of the 1% egg albumin solution and pour it into a test tube. Add 1 ml of dilute
sodium hydroxide solution and swirl to mix. Using the dropper, add a few drops of copper
sulphate solution and swirl again. Record all observations.

TEST FOR REDUCING SUGARS

To 1ml of 1% glucose in a test tube, add 1ml of Benedict’s reagent and swirl. Place the test tube
in a water bath for a few minutes and record all observations.

TEST FOR NON-REDUCING SUGARS

To 1ml of sucrose solution in a test tube, add 2 drops of HCl and swirl. Place the test tube in a
water bath for 2 minutes. Allow to cool and add 1ml of NaOH solution and swirl. Add 1ml of
Benedict’s reagent and swirl. Place the test tube in a water bath for a few minutes and record
all observations.

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FOOD SAMPLE FOOD TEST OBSERVATIONS INFERENCES

1% Starch Starch

1% Glucose Reducing
Sugar

1% Sucrose Non-reducing
Sugar

Oil Lipids

1% Egg Starch
Albumin

Reducing
Sugar

Non-reducing
Sugar

Protein

Lipids

Chicken Stock Starch

Reducing
Sugar

Non-reducing
Sugar

Protein

Lipids

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Or Alternatively: June 2009, Paper 3/2 Question 1

1. Amylase is an enzyme, which catalyses the hydrolysis of starch into a reducing sugar. It is
commonly found in human saliva and germinating seeds. You are required to carry out a
simple investigation into the effect of substrate concentrations on the rate of reaction of
amylase. Carefully read the instructions that follow before starting.

(a) You are provided with a stock solution of starch. Using the distilled water provided prepare
a series of starch solutions at different concentrations as follows:

(i) Label 8 test tubes A, B, C, D, A l , Bl , Cl , D1

(ii) To EACH test tube add the following:

A: 5 cm3 stock starch solution

B: 4 cm3 stock starch solution and 1 cm3 distilled water

C: 3 cm3 stock starch solution and 2 cm3 distilled water

D: 2 cm3 stock starch solution and 3 cm3 distilled water

A1 , B1 , C1 , Dl: 5 cm3 distilled water

(b) Place all the test tubes in a test tube rack in the water bath (37 °C) provided and leave for 5
minutes to allow the temperature of the solutions to reach the temperature of the water
bath.

(c) Remove test tubes A and Al from the water bath. To each tube quickly add 0.5 cm3 of the
amylase enzyme solution provided. Swirl the contents of the test tubes and rapidly put them
back into the rack in the water bath.

(d) START THE STOP CLOCK. At intervals of 1 minute remove 1 drop of the reaction mixture
from each test tube and place in separate wells of the marble spot plate provided.
Immediately add a drop of Benedict's solution. Record the starch concentration and colour
changes in Table 1 on page 3. Repeat this for a period of 5 minutes.

(e) Carry out the same procedure outlined in Steps (c) and (d) for the remaining pairs of
solutions as follows :

B and B1
C and C1
D and Dl
(f) Write an appropriate title for Table 1.

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TABLE 1 :______________________________________________________________________

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(g) Based on the results recorded what can you deduce about the relationship between
substrate concentration and enzyme activity?

(h) Comment on the purpose of the distilled water in test tubes A 1 to D 1.

(i) Give TWO reasons for the colour changes recorded over the five-minute test period, when
Benedict's solution was added to the reaction mixtures.

(j) Outline the procedure that you might use to produce quantitative colour standards for the
reaction mixtures using Benedict's solution.

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4. Quantitative Investigation and Comparison of Reducing Sugars and Starch

Refer to Plan and Design item (ii) on page 32

5. Diagrams and Electron Micrographs of Cell Membranes and Organelles in

Animal and Plants

AIM: To make diagrams of cell membranes and organelles in animal and plant cells from
electron micrographs

APPARATUS AND MATERIALS:

Projector, laptop, powerpoint entitled: Electron micrographs of cell membranes and organelles
in animal and plant cells

METHOD:

Make detailed diagrams of each micrograph presented.

6. Electron Micrographs and Prepared slides of Prokaryotic Cells

AIM: To make diagrams of prokaryotic cells from electron micrographs and prepared slides

APPARATUS AND MATERIALS:

Microscope, prepared slides of prokaryotes, projector, laptop, powerpoint entitled: Electron

micrographs of prokaryotic cells

METHOD:

Make detailed diagrams of each micrograph presented.

27
7. Tissue Plan and Detailed Diagrams of Tissues in Dicotyledonous Plant
Roots and Stems

AIM: To make detailed diagrams and tissue plan diagrams of tissues in dicotyledonous stems
and roots from prepared slides

APPARATUS AND MATERIALS:

Prepared slides of dicotyledonous stems and roots, microscope

METHOD:

Make detailed diagrams and tissue plan diagrams of each specimen.

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8. Investigating the Effect of Immersion into Solutions of Differing Water
Potentials on Plant Cells

AIM: To investigate the effect of solute concentration on Ψ in plant cells

Using the 0.1M Sucrose stock solution provided, make up 20ml of each of the following
concentrations: 0.02, 0.04, 0.06, 0.08, 0.1 M. Pour each solution into labelled petri dishes and
cover.

Select a large firm potato tuber (Solanum tuberosum) and cut out 10 cylinders using a 5-6mm
diameter cork borer. Trim each potato cylinder to 40mm in length using a razor blade making
sure that all potato skin is removed. Place each cylinder immediately into a small beaker that is
kept covered until all the cylinders are cut. Measure all cylinders to ensure that the length is
exact to the nearest 0.5mm and place two measured cylinders into each labelled petri dish.
Note the time and cover each petri dish. Leave the dishes to stand for exactly 60 minutes then
remove the potato cylinders from each solution. Blot lightly on a paper towel and quickly
measure the new length of each potato cylinder to the nearest 0.5mm. Please record all
measurements and observations in an appropriate format.

Please show all working. You will be required to calculate the following:

1. How much of the 0.1M sucrose stock solution is needed to make up 20ml of each
solution concentration identified.
2. The average change in length of each potato cylinder. Use + or – to indicate an increase
or decrease in cylinder length respectively.
3. The percentage change in length of each potato cylinder in each solution.
4. Plot a graph of change in length against sucrose concentration.
5. Determine which sucrose concentration results in zero change in cylinder length.
6. Determine the water potential of the potato tissue.

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9. Effect of Temperature on Enzyme Reactions

AIM: To investigate the effect of temperature on enzyme catalysed reactions

APPARATUS AND MATERIALS:

Stop watch, test tube holder, test tubes, test tube rack, 400ml beaker, 200ml beaker, 25ml
measuring cylinder, thermometer, rubber bung, delivery tube, stirring rod, distilled water,
Bunsen burner, tripod stand, wire gauze, delivery tube, boiling tube and Solution S1(mixture of
yeast, water, glucose)

METHOD:

Please READ your procedure before coming to the lab session to ensure that the enzymes are
not denatured before the experiment starts. Identify any precautions you might need to take.

Step 1: Prepare a water bath using the 250ml beaker and maintain a temperature between 25
and 300C. Label 2 boiling tubes T1 and T2.

Step 2: Add 20ml of S1 to T1 and fit the rubber bung and the delivery tube. Add 20ml of tap
water to T2 and fit the rubber bung and delivery tube. The other end of each delivery
tube is to go into separate test tubes of cold water.

Step 3: Place the boiling tube end in the water bath and allow S1 to equilibrate for 3 minutes.
Count the number of bubbles generated in 30 second intervals for 2 minutes for both T1
and T2 at this temperature. Record results in a table.

Step 4: Increase water bath temperature to a range between 35 and 40 0C and repeat step 3.

Step 5: Increase water bath temperature to a range between 45 and 50 0C and repeat step 3.

RESULTS:

Please tabulate your results and plot a graph in the form of a bell shaped curve.

DISCUSSION:

Please discuss your results fully and identify any limitations and sources of errors.

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10. Effect of Substrate Concentration on Enzyme Reactions

AIM: To investigate the effect of substrate concentration on enzyme catalysed reactions

APPARATUS AND MATERIALS:

Stop watch, test tube holder, test tubes, test tube rack, 10ml measuring cylinder, rubber bung,
delivery tube, stirring rod, boiling tube, scalpel, ruler, potato cylinder, white tile, labels, small
beaker, dropper and 2M hydrogen peroxide solution.

METHOD:

Please READ your procedure and complete ALL calculations for the hydrogen peroxide dilutions
before coming to the lab session. Identify any precautions you might need to take.

Step 1: Place 10ml of 2M hydrogen peroxide solution into a boiling tube. Label it 1 and replace
rubber bung with delivery tube. Label a test tube 2 and add enough tap water to it so
that the level is 1cm higher than the end of the delivery tube. NOTE: If it is too low your
bubbles will be lost.

Step 2: Place a potato cylinder on the white tile and line up the ruler alongside it. Cut 10 discs
2mm wide from the cylinder with your scalpel. Put the discs into tube 1 and gently shake
by swirling to separate the discs. Replace rubber bung with the delivery tube in tube 2.

Step 3: Wait for 30 seconds then count the number of bubbles evolved in one minute for
3minutes. Record each value then calculate the average for that hydrogen peroxide
concentration.

Step 4: Make up 10ml of a 1.5M dilution of the hydrogen peroxide and repeat steps 1 to 3.

Step 5: Repeat steps 1 to 4 with a 1M and 0.5M dilution of hydrogen peroxide.

RESULTS:

Please tabulate your results and plot a graph in the form of a S-shaped curve.

DISCUSSION:

Please discuss your results fully and identify any limitations and sources of errors.

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11. Stages of the Cell Cycle from a Root Tip Squash and Prepared Slides

AIM: To make detailed diagrams of the stages of the cell cycle from prepared slides of a root tip
squash and show the stages of mitosis from a fresh root tip squash

APPARATUS AND MATERIALS:

Prepared slides of the Allium root tip squash, onion root, orcein acetic, slide, coverslip, tissue,
distilled water, dropper

METHOD:

Make detailed diagrams showing the stages of the cell cycle from the prepared slide. Prepare a
wet mount of 1 onion root using orcein acetic stain. Gently press the root under the coverslip to
squash the specimen without breaking the coverslip. Make detailed diagrams of the cell cycle.

12. Meiosis Model

AIM: To construct models to demonstrate chromosome behaviour in meiosis

APPARATUS AND MATERIALS:

You may use any materials you want to for example: pipe cleaners, thread, wool, twist ties,
beads, Bristol board.

METHOD:

Using the materials you identified, construct models to demonstrate chromosome behaviour in
meiosis. Ensure that the items selected are securely fastened to the backing material you
select. A short note on each stage should accompany each stage identified and constructed in
the models.

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13. Stages of Meiosis from Prepared Slides of the Testis or Anther

AIM: To make detailed diagrams of the stages of meiosis in the testis or anther from prepared
slides

APPARATUS AND MATERIALS:

Prepared slides of testis or anther, microscope

METHOD:

Make detailed diagrams and tissue plan diagrams of the specimen.

14. Drawing of the Anther and a Plant Embryo Sac

AIM: To make detailed diagrams and tissue plan diagrams of tissues in the anther and a plant
embryo sac from prepared slides

APPARATUS AND MATERIALS:

Prepared slides of the anther and a plant embryo sac, microscope

METHOD:

Make detailed diagrams and tissue plan diagrams of each specimen.

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15. Seed Samples and Mendelian Genetics

PART 1:

AIM: To investigate a dihybrid cross in an ear of corn.

METHOD: The four different genes and grain types are identified in the following photo:

There are four grain phenotypes in the above ear of genetic corn: Purple & Smooth (A),
Purple & Shrunken (B), Yellow & Smooth (C) and Yellow & Shrunken (D). These four grain
phenotypes are produced by the following two pairs of heterozygous genes (P & p and S &
s) located on two pairs of homologous chromosomes (each gene on a separate
chromosome):

DOMINANT GENES RECESSIVE GENES

P = Purple p = Yellow

S = Smooth s = Shrunken
A dihybrid cross between two heterozygous parents of the genotype PpSs was done producing
the ear of genetic corn shown below.

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Identify the 4 gametes of each parent and write the four gametes of each parent along the top
and left sides of the table. Write an appropriate title for Table 1. Complete the Punnet square
shown in Table 1 to show the dihybrid cross between the parents.

TABLE 1: _____________________________________________________________________

GAMETES

Questions:

1. What is your dihybrid ratio?

2. Is your ratio the same as the expected ratio?

R.C Punnett, a colleague of William Bateson devised this method. In 1900, English Geneticist
William Bateson had Gregor Mendel's original 1865 paper on the genetics of garden peas
translated into English and published and Mendel became known to the entire scientific world.
Bateson is also credited with the discovery of gene linkage in 1905.

Calculate the number of different phenotypes and genotypes in a dihybrid cross using the
following formulae: Number of phenotypes: 22 = 4
Number of genotypes: 32 = 9

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PART 2:

AIM: To Conduct a Chi Square test to determine if the data provided follows the theoretical
dihybrid ratio.

An ear of corn has a total of 381 grains, including 216 Purple & Smooth, 79 Purple & Shrunken,
65 Yellow & Smooth, and 21 Yellow & Shrunken.

State your hypothesis and use the chi square test and probability values to test your hypothesis.
In order to test your hypothesis you must fill in the columns in the following Table 2.

Write an appropriate title for Table 2 below.

TABLE 2: _____________________________________________________________________

Grain Observed Observed Expected Expected [Obs No. - Exp

Phenotype Number Ratio Ratio Number No.]2
÷ Expected No.

Purple &
Smooth
Purple &
Shrunken
Yellow &
Smooth
Yellow &
Shrunken
Total
Number:

Chi Square Value:

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16. Drawing of a Mammalian Testis and Ovary

AIM: To make detailed diagrams and tissue plan diagrams of tissues in dicotyledonous stems
and roots from prepared slides

APPARATUS AND MATERIALS:

Prepared slides of dicotyledonous stems and roots, microscope

METHOD:

Make detailed diagrams and tissue plan diagrams of each specimen.

17. Planning and Designs

ii) Plan and design an experiment to investigate the permeability of visking tubing
iii) Plan and design an experiment to investigate the sugar content in ripening fruit
iv) Plan and design an experiment to investigate the effect of substrate concentration on
enzyme activity
v) Plan and design an experiment to investigate the activity of amylase at different enzyme
concentrations
vi) Plan and design an experiment to investigate the action of amylase on starch
vii) Plan and design an experiment to investigate the amount of sugar present in sugary
drinks consumed at school with the amount present in those consumed outside of school

18. The Investigative Project

The nature of the Project will be disclosed at a later date.

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