1

Unit 3 Basic Principles

❖ Outline of Any Experiment:
1- Independent variable: the factor you change ex. temperature:
2- Dependent variable: the factor you measure
ex. rate of the reaction /mass of product. ( always include SI unit)
3- Controlled variables kept constant.(for / ensures* Validity)
(not increase* validity) . They are factors that can affect your results.
-You need to give ( whenever possible)
a--Examples of controlled variables. (+/- suggested values).
b--How to control (apparatus).
c--Effect on the results if not controlled ( obligatory in unit 6 ).
-Mention controlled variables in:
a- organism: (same age , species ,mass,gender)
b- experiment: ex. same pH, volume and concentration of (enzyme).
c- surrounding conditions: (temperature, light intensity)
d- others : same type and mass of food provided to an organism.
4-Time scale : suitable to show the expected results.
5- Replications: Minimum 3 times at EACH….
(For reliability, NOT to increase xxx reliability)!
-(to spot anomalies , minimise the effect of individual variations,)
-to calculate the mean , show variability, SD, and allow statistical analysis).
6- Control: , Definition: Identical experiment, with identical apparatus and
conditions, without the factor you are testing (ex. without enzyme).
- Purpose of the control: Allows comparison –( Ensures* VALIDITY) to
be sure that your results are really due to this factor & no other factor.

1
 2

e.g.. Experiment result = 50 au ,, control result = zero au

Actual result of experiment is: 50 – zero = 50 au.
e.g. Experiment result = 50 au. Control result = 4 au.
Actual result of experiment is: 50 – 4 = 46 au.
❖ Validity : How TRUE/correct your results are?? This is achieved by:
1- Keep all controlled variables constant.
2- Use a control to compare.
3- Choose the correct accurate instrument to measure your parameter.
4- Non overlapping error bars→ SIGNIFICANT difference /correlation.
❖ Reliability: ( Can you trust the results) ?
1- Large sample size ( individuals / repetitions) to be representative.
Also anomalies can be identified and excluded.
2- Low Variability: Consistent /similar/ close results of repetitions,
/clustered around the mean → Small SD / narrow range / error bars.
3- Non overlapping error bars→ SIGNIFICANT difference or correlation.
4- Repeatable by other scientists, giving the same/similar results.
NB: (repeating does NOT make your measurements more reliable, it just
helps you to spot and remove anomalies, asses and judge the degree of
variability, hence reliability!)
• Random error: Unpredictable , occasional error in one* or more of the
measurements, usually caused by:
1- problem in the controlled variables , or
2 -inaccurate instrument.
-Ex. when weighing yourself, and standing differently on the scale,
or looking at the level of a liquid each time from a different angle, or
difficulty in standardising the pressure or force applied.
-Corrected by : Performing many repetitions, exclude the anomalies.

2
 3

• Systematic error: persistent error, always obtained, with the same , due
to poorly calibrated apparatus
- Corrected by: proper calibration of your apparatus, during the planning
stage, or preliminary work.
[ If a non-calibrated balance gives an extra 10 grams mass each time you
use it , ( reading is always 100 grams, while the real is 90 grams)→
(systematic error). This means your results are not accurate!!,
❖ Accurate: your measured results are close to the real/ actual/ standard
values. -How to improve accuracy :
1-Use a more sensitive instrument, and repetitions to minimize random
errors.
2-Calibrate your apparatus ( to remove systematic error).

-Qualitative result: describing what is present / not present. ex judging the

colour by the eye ex biuret test, -ve = blue, +ve purple
-Semi-Quantitative result:
ex. comparing the colour obtained to a colour chart.
Disadvantage of both of the above : subjective , inaccurate,
-Quantitative result: numerical/digital, accurate,
( result indicates exactly how much is present)
ex by using a colorimeter..
Advantage : objective, more accurate , can be used to draw a graph
and do statistical analysis.

3
 4

Conducting an Investigation

Preliminary work( Planning ):

1-Try the proposed method to see if it will work

2-To decide upon the best factors that would give meaningful results:
SELECT: Most suitable/best:

1- Organism: Type/species, source, age, ….

2- Independent variable: The most suitable range and intervals.
3- Dependent variable: (What will you measure?) Best parameter
e.g. mass, length for growth.
4- Apparatus: Choose the correct, most accurate, calibrated apparatus.
5- Method: Try different methods to see which works best.
6- Set most important controlled variables to ensure validity.
7- Consider the possible risks and safety precautions, ethical concerns.

• Ethical concerns: Explain clearly:

1- Consider the welfare of animals used, and the concept of animal rights
in vertebrates and invertebrates.
2- Ethical concerns of harming or disturbing the surrounding environment.
Subjects should give consent to participate.
3- Otherwise say: { There is no significant ethical concerns }

4
 5

A) Risks and Safety precautions (just examples)

Risk How to minimise it

Injury when cutting with a scalpel Cut away from one’s self , wear gloves
or knife and from broken
glassware.
Infection from bacterial cultures Use sterile equipment,,, see the experiment

Allergies, irritation, corrosion from Wear protective gloves, masks, goggles

chemicals e.g. enzymes, stains ….
Risk of fire from flammable Handle flammable substances away from
substances e.g. alcohol used near any fire or Bunsen burner, wear heat
Bunsen burner flame. resistant gloves. Heat indirectly, use electric
plate, water bath.

Splashes from heating liquids or Wear goggles

specs of broken fibres hitting the
eyes.
Insect bites or falling branches in Wear protective clothes
field ecological sampling studies
Harm from handling animals Wear protective gloves ,eye protection

(Go back to each Individual experiment for more safety precautions)

5
 6

B) Method: (Clear and detailed)

1- Identify ( mention) the independent variable : clearly.

2- State 5 specific values,(should be in the range and intervals to give

meaningful results) / or
-the presence and absence of the IV variable.

-Put your units clearly and apparatus used to set the values
(ex. incubator for temperature)

3- Identify the dependent variable: ( must be a measurable parameter,)

(ex dry mass/Length to measure growth )

State the apparatus used to measure it. -Use SI units.

5- Use a control to compare ( in most experiments ).

6- Describe your method briefly

-Mention and justify the tool or instrument used to measure the

dependent variable .

-Indicate any calibration if needed, ex that of colorimeter, or spirometer.

7 - Time scale: Suitable to show the results

8- Controlled variables ( in the organism, conditions,apparatus)

- Mention at least two ( in the organism, + chemicals used ).

- Put suitable values if possible.

- Mention how each is achieved ( at least one)

9- Replications: several times at EACH…. and used to get average.

reasonable number to ensure reliability ( too few →non-representative,
too many → not practical + time consuming)

6
 7

C) Recording, presenting and analysing the data:

➢ Recording : in clear tables with SI units.

➢ Presenting: GRAPH
➢ Analysing : Name + justify (reason) suitable statistical test.
-t- test: for difference between 2 or more groups ( independent variable
is discontinuous, ex. in males versus females , or in different countries)

- Spearman rank correlation: when the independent variable is

continuous, ex, increasing light intensity.

- Chi – square test for the difference between the observed and
expected (ex in genetic crosses).
-Interpretation: In all of them: If the calculated value is greater than the
critical value at 0.05 significance level, -→ then there
IS A SIGNIFICANT DIFFERENCE / CORRELATION!
-The closer the correlation coefficient to 1, the stronger the correlation.

D) Limitations (Mostly unavoidable!): .--> affects the validity!

❖ Difficulty in controlling all the variables: ex.
- Light intensity a field (a cloud passing).
- Genetic variation in organisms.
- Antibiotic molecules/ chemicals may have different sizes, so different
rates of diffusion through the agar.
❖ Difficulty in determining the end point or measuring the dependent
variable e.g. colour change in DCPIP.
❖ Cells overlapped in root tip squash.
❖ Behaviour of organisms in the lab may be different from real life.
❖ Difficulty in identifying the species when similar ones exist.

7
 8

GRAPHS

Axes: Correct orientation (choice):

• Independent variable on X axis (that you change).
• Dependent variable on Y axis (that you measure).
Labelling: Very clearly with SI units!!! ( IN BRACKETS )
Type of graph: - Bar , it the IV is discontinuous.
- Line , if the IV is continuous.
Scale: Take care!!
1- Use more than 50% (1/2) x axis , and also more* than 50% (1/2) y axis.
2- If all your readings are within a narrow range of large numbers e.g. 201,
203, 207, 209, you can interrupt ( split / Truncate) the axis from the zero,
or start from 200.( However, You Cannot do that in BAR graphs!!!)
3- SD or RANGE ..VIMP!!!, to be considered when deciding your scale,
(maximum value , see sample old syllabus!)
Plotting the data: Rules:
1- Use clear dots / crosses ± circles. If bars are used = equal width !.
2- If 2 or more sets of data = use a Key and different symbols / shades.
3- Never go out of the graph paper (white area).
4- Never extrapolate forwards or backwards !! (you lose one mark if you do)
[Never assume that at zero X, the value of Y was zero !]
5-The points joined clearly by broken line ( ruler !) (not free hand)!
6- Don't use line of best fit unless you are asked to do so.
( Fairly equal points on either side of the line).
• Error bars (variability) using S.D. or Range ,
Only plotted if the examiners asks for it ! (don't volunteer) (too much time).

8
 9

Describing graphs:
❖ Graph 1
M
Risk of CVD (au)

Smoking level (au)

➢ Trend, Steepness, Manipulation

1] Describe the relation between smoking level & CVD risk in males and females
1-In Both males and females, as smoking level increases ,the risk of CVD increases.
(+ve correlation).
2- Linear increase in both.
3-Risk in males is always higher than females .
4-Steeper increase in males than in females.
5-Manipulation: Subtract, ratio , % (choose the easiest). not just copy figures !
ex. At smoking level 2 au, the risk in males is higher than females by 4 au. ( never
forget the unit!!!) fixed the 2 au on the X axis ,and subtracted 8 - 4 on the Y axis
-wrong: At smoking level 2 au, the risk in males is 8 au while that in females is 4 au
-wrong: the risk in males by - - - - XX (because you didn’t mention females) !!
2] Give evidence from the graph that there are other factors/ genetic factors
linked to increased risk of CVD -Even non smokers (smoking level zero!) have some
risk of CVD in both males & females.
9
 10

❖ Graph 2
In this graph, points of trend and manipulation are
Risk
the same as graph 1, however, since both lines are
parallel, they have same steepness !! (point 4 )

Smoking level

Error Bars (variability)

Show the degree of spread of the results of the repetitions around the mean.
• Error bars can be obtained by:
a) Range: simply difference between highest & lowest results of the repetitions.
(However, one anomaly would give a wide error bar based on the “Range”.,,,so
better use SD→
b) Standard deviation: S.D. shows the average degree of spread of the data around
the mean, considering the sample size ( The number of repetitions.)
It is calculated by a certain equation ( later). This is more objective, as not greatly
affected by only one or two anomalies within large number of repetitions.

S.D. of the same

Range experiment &
same results

10
 11

A) Overlap of error bars ( significance)

- When assessing whether there is /is not a “ SIGNIFICANT
difference/correlation” between groups ex. blood pressure with and without
using a certain drug.
overlap

No
overlap

   

- No overlap of error bars - Overlap of error bars

- No common values bet. gp 1 & gp 2 - Common values bet. gp 1 & gp 2
- The highest values of gp are
- similar to the lowest values of gp2
- There is a significant difference - Non* significant difference
between the results of the 2 groups.
- You can make a valid conclusion→ - May be due to chance.
This drug may be effective. -

Confirm by statistical tests in A2 !!

11
 12

B) Size of error bars (variabilty)

-Narrow error bars -Wide error bars
-Low variability, -High variability,
→ - High reliability → - Low reliability

mean 10 mean 10

(A) (B)

The mean of both groups can be the same, but in (A), the
results of repetitions are more similar/close/clustered to the
mean, → lower variablity so more reliable!!

Tabulation:
1- Independent variable → first column.
2- Dependent variable in the second.
3-Units in headings only* , between brackets ,use SI units.
4-Include columns for the repetitions if required, and the mean.
5- Use horizontal lines to separate the data.
6- Use consistent number of decimal places. Ex if 2.87 in the data, the
zero has to be 0.00.
7-
12
 13

❖ Diluting a stock solution:

Making serial dilutions: With a dilution factor 2:

1- Add a certain volume from the original stock solution (ex 5 cm3)
2- Add EQUAL volume of distilled water ( also 5 cm3).
3-Total volume would be 10 cm3.
4- Do this repeatedly,,,Ex. obtained concentrations would be :
100%, 50%, 25%, 12.5%, 6,25%.....

𝒕𝒐𝒕𝒂𝒍 𝒗𝒐𝒍𝒖𝒎𝒆
• Dilution factor =
𝒗𝒐𝒍𝒖𝒎𝒆 𝒐𝒇 𝒔𝒕𝒐𝒄𝒌 𝒕𝒓𝒂𝒏𝒔𝒇𝒆𝒓𝒆𝒅
10
=2
5

13
 14

What would be the dilution factor be in the examples below?

14
 15

Q) What is meant by molar solution of glucose?

- It is the molecular weight of glucose in grams dissolved in distilled water and
made up to [ 1 dm³ ].
Q) How can you prepare serial dilutions from a stock of glucose solution of
concentration 1 mol /dm³?

Apply the formula C₁ V₁ = C₂ V₂

C₁ is the concentration of stock solution
V₁ is the volume you are going to take from the Stock to be diluted.
C₂ is the concentration of the solution you are going to prepare.
V₂ is the volume of the solution you are going to prepare.
Suppose you want to prepare 10 cm³ with the following dilutions respectively

0.5, 0.25, and 0.1 mol dm̄ ³ from the stock (1 mol dm-3)

A)for first dilution (0.5 mol dm-3)

C₁ V₁ = C₂ V₂
1 X V1 = 0.5 X 10
0.5 𝑥 10
V₁ = = 5 cm-3 from the stock, then make them up to 10 cm³ by distilled
1
water.
B)For second dilution [0.25 mol dm̄ -3),
C₁ V₁ = C₂ V₂
1 x V₁ = 0.25 x 10
V₁ = 2.5 cm-3 from the stock, then make them up to 10 cm³ by distilled water.
C)For third dilution [0.1 mol dm-3]
C₁V₁ = C₂ V₂
1 x V₁ = 0.1 x 10
V₁ = 1 cm³ from the stock, then make them up to 10 cm³ by distilled water

15
 16

Questions:
1. Describe how you would make 50 cm³ of a glucose solution of concentration
0.05 mol dm̄ ³ from a glucose solution of 0.1 mol dm-3 ?
C₁ V₁ = C₂ V₂
0.1 x V₁ = 0.05 X 50
0.05 𝑥 50
V₁ = = 25 cm³
0.1
-Measure out 25 cm³ from the stock [0.1 mol dm-3], make up to 50 cm³ by adding
25 cm³ distilled water.
2. Starting with stock solutions of 10% glucose and 2% sucrose how would you
make 100 cm³ of a mixture of final concentration of 1% sucrose and 1% glucose?
Stock glucose solution: C₁ V₁ = C₂ V₂
10 x ? = 1 x 100
𝟏𝟎𝟎𝒙𝟏
V₁ = = 10 cm³ from stock glucose solution
𝟏𝟎
Stock sucrose solution: C₁ V₁ = C₂ V₂
2 x V₁= 1 x 100
𝟏𝟎𝟎𝒙𝟏
V₁ = = 50 cm³ stock sucrose solution.
𝟐

Add 10 cm³ of 10% glucose to 50 cm³ of 2% sucrose and make up to 100

cm³ with distilled water.

16