CAIE A2 LEVEL BIOLOGY (9700)

Statistics Criteria Formulae Interpretation

1. Data Analysis 2 sets of χ2 = *v ∗ =

x2 − test discrete/nominal (O−E) 2 (∗n ∗ −
data ∑ E 1)2
​ ​

1.1. Measures of Central Tendency, null and

h0 = h1 = X 2 >
Location & Dispersion alternate
​ ​

X 2 < CV CV
hypothesis →
Mean is sum of data divided by no. of data.
2 sets of normal,
Mode is most common value. -1 : (-) relation
Pearson’s discrete r=
Median is middle quartile. \n 0 : no
linear quantitative ∑ xy−n(x)(y )
Range is spread between smallest and largest value. relation \n + 1
​ ​

correlation data(>5 n(sx )(sy )

​

: (+) relation
​ ​

It can be divided into 4 quarters by 3 quartiles.

readings)
Interquartile range is spread between upper and lower
quartile.
2 sets of
Spearman’s r0 = 1 − 0 : no relation
discrete/ordinal, ​

∑ (x − x) 2
rank \n 1: true
( )
2
Standard deviation (s) = 6− ∑ D
normal data (10-
​

n−1
correlation relation
​
​

n3 −n
​

Standard deviation is spread of data around mean. Data 30 readings)

with a narrow spread is more reliable than that with a Simpson’s
wider spread.
D = 1 − 0 : not diverse
index of Population data 2 \n 1 : very
Standard error (sm ) = sn
​ ​

diversity
∑ ( Nn ) diverse
​ ​

Standard error measures the reliability (of the estimate) n=

Mark-release- n = total
of a mean. The larger the standard error the less reliable. Population data n1 ×n2
capture population
​ ​

n (marked)
​

Confidence limits are the ranges/intervals in which the

true value of the mean lies with 95% probability. 95%
confidence intervals (represented graphically by error
bars) lie within 2 standard deviations/errors of the mean.
2. Common Investigations

1.2. Statistical Tests & Calculation 2.1. To investigate the effects of salt
solution of different concentration on
Statistically significant: it is an idea that the (observed)
results are caused by an outside potato strips
factor (not due to chance).
P < 0.05: it is an idea that 0.05 means that there is less Make at least 5 new salt solutions (of 5 different
than 5% chance of obtaining the (observed) results by concentrations) by diluting (serial or simples) the stock
chance. salt solution
Bar chart shows Keep volume of all the solution same
discontinuous/discrete/qualitative/categoric data. Measure volume using graduated
Chi square test is used for data which is pipette/burette/measuring cylinder
categoric/discrete. Repeat experiment at least three times with each solution
Pearson’s linear correlation test is suitable when (both and take mean
sets of) data are continuous and (are approximately) Use same number/size/mass of strips in each solution
normally distributed; a scatter graph or data suggests a Measure strips using callipers/ruler
linear correlation. 5 or more paired observations are Measure mass with electronic balance
required to carry out this test. Use strips from the same potato
Keep strips in solution for the same amount of time
Statistics Criteria Formulae Interpretation Measure time using stopwatch
2 sets of normal, t=
continuous ∥x− y∥ v = nx + 2.2. To investigate the effects of
t-test
​

quantitative data s2x ​

s2
+ nyy
​
ny − 2
​

different concentration of enzymes on

(>5 readings) nx
​ ​

​ ​

null and the activity if enzymes

h0 = t < h1 = t >
alternate
​ ​

CV CV Make at least 5 new enzyme solutions (of 5 different

hypothesis →
concentrations) by diluting (serial or simple) the stock
enzyme solution
Keep volume of all the solution same

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CAIE A2 LEVEL BIOLOGY (9700)

Measure volume using graduated Calculate volume by measuring diameter capillary and
pipette/burette/measuring cylinder multiplying by the length/distance moved
Repeat experiment at least three times with each solution
and take mean 2.5. To investigate the effects of carbon
Keep pH value most suitable for enzyme to work (add
acid/alkali/buffer to keep pH constant) dioxide concentration on the rate of
Incubate enzyme at suitable working temp of enzyme photosynthesis
(incubator, thermostatically controlled water bath)
Use same size/shape of sample/strips on which the
enzyme is acting
Continue experiments for the same period of time
Immerse sample/strip and start stopwatch
simultaneously

2.3. To investigate the effects of

different concentration of solution on
the number of cells being plasmolysed
Cut strips of sample using tweezers/scalpel
Standardize sizes of strips (measure length using
callipers/ruler mass using electronic balance)
Use hydrogen carbonate to provide carbon dioxide
Same number of strips in all different concentrations Make at least 5 different concentration of carbon dioxide
Leave for same period of time Measure concentration of carbon dioxide with a probe
Mount on slide in sucrose solution (with coverslip); (see Measure the time taken to collect a known volume of gas
slide preparation technique) Use same mass/length/same piece/same number of
Count large number of cells, count more than once and in
leaves/same species of pondweed
more than one field view, take mean Keep light at a fixed distance from plant (light intensity)
Record number of plasmolysed and non-plasmolysed Use thermostatically controlled water bath
cells
Use same volume of water (measuring cylinder, burette)
Carry out experiment in a dark room (eliminate any other
2.4. To investigate the respiration rates light source)
Use 3 sets of measurements per experiment and take
of small invertebrates and germinating mean
seeds using respirometer Low risk experiment (electric shocks faulty
equipment/wet wiring)

2.6. To investigate transpiration in

plants

Measure mass of invertebrates and germinating seedling

Use seedlings before plumule emerged
Dye at capillary tube farthest from tube
Keep Airtight using the air-tight bung
Keep temperature constant
Allow organism to adjust for a given period of time
Close clip before taking measurements
Measure distance moved by dye for standard amount of
time
Repeat experiment at least 3 times
Replace carbon dioxide absorbent (e.g. Soda lime)
between measurements

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Use shoots/leaves of different types of plant used

Keep surface area of shoots/leaves same
Measure/record the movement (of water) along the
capillary
Measure time of water movement for a fixed amount of
distance using stopwatch
Keep light intensity, temperature, and humidity same
Carry out experiment in a dark room with light of fixed
illumination
Keep temperature constant e.g. temperature-controlled
room
Insert shoot under water/cut shoot at an angle
Dry the leaves before measuring mass
Keep airtight using an airtight apparatus/rubber bang
Observe dye through stem, at known time interval and Use a syringe to set water level in capillary
known distance; record time for dye to reach it Leave until equilibrium reached
Use Vernier callipers to measure stem and ruler in Repeat experiment for at least 3 times and take mean
cm/mm
Use several shoots/sequential measurements on the 2.8. To investigate the concentration of
same shoot
Cut under water/dye to avoid air from entering
enzymes in a given extract (e.g. starch
Keep temperature constant e.g., temperature-controlled agar well)
room
Carry out experiment in a dark room with light at fixed Dilute the stock enzyme solution using serial dilution to
illumination make at least 5 solutions of different concentrations
Keep light at a fixed distance Use denatured enzyme as control
Provide steady air flow using a fan set at constant speed Use ruler / callipers / string and ruler to measure
Repeat experiment for at least 3 times and take mean diameter of the well
Low risk experiment: dye can be toxic, wear gloves Plot a calibration curve of known concentrations and use
it to determine extract concentration
2.7. To investigate the transpiration Use the same volume of enzyme
Leave all plates for the same period of time
rates of two different plants using a Use a thermostatically controlled water bath to keep
potometer temperature constant
Use a buffer to keep the pH of the agar same
Use same concentration of starch in the agar plates
Keep same depth/volume of agar in Petri dish
Cover to prevent contamination / evaporation
Repeat experiment at least 3 times and calculate mean
Low risk investigation: agar is irritant and wearing gloves

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medication; dangerous environment; hazardous

2.9. To investigate the change in
plants/hazardous animals; wearing suitable shoes
distribution and abundance of the plant
species as the distance from the edge 2.11. To investigate the effect of
of a pond is increased using systemic caffeine/addictive substances heart
sampling method rate/pulse rate/reaction time in
animal’s (e.g. Daphnia) body
Use belt (interrupted or continuous) or line transect
Use measuring tapes with marks to measure distance / Use a large number of test subjects
length, of transect Select where around pond/river to
Provide groups with drinks with and without
place the
caffeine/addictive substance
transect
Allow animals to acclimatize after adding substance
Use frame quadrat/point frame/point quadrat Use a clicker/tally counter to count the number of heart
Sample continuously at regular intervals beats
Use same/stated size, quadrat/frame/point
Count for the same period of time
frame/sample area
Use same volume/same number of drops of substance
Identify different species using a guidebook/photograph
solution added to each drink
Estimate distribution by counting/calculating
Making drinks indistinguishable
percentage/density) Make sure test subjects do not take in any substance for
Replicate transect at least once
at least 5 hours before the test
Sample at different times of year/seasons
Test each subject in isolation/away from others
Safety: injury/getting lost and staying with a group; allergy
throughout the experiment
to plants and wearing gloves/protective clothing; allergy Make sure subject is at rest whilst having measurements
to pollen/hay fever and wearing mask or taking (reaction time/heart rate)
medication; dangerous environment; hazardous
Take measurements (of reaction time/heart rate) before
plants/hazardous animals; wearing suitable shoes
giving the drink
Wait a minimum of 45 minutes after giving the
2.10. To investigate a random drink before measuring the reaction time and heart
rate
(unbiased) method to collect data
Give the same volume of drink to all subjects
needed to calculate the biodiversity of Use subjects of same age/mass/weight/fitness
level/ethnicity/race
plant species
Standardize sex balance (equal number of one sex)
Calculate mean for the measurements
Mark out the area to be sampled using tape
measures/string and marker pole/ Make sure that subjects have no pre-existing medical
Generate random number (for coordinates) using a conditions that may affect the experiment
mobile application Allow test-subjects to stop if they feel unwell
Make a minimum of three replicates and calculate mean
Use a frame or point quadrat for individual samples
Place quadrat at the coordinates 2.12. To investigate how the rate of
Identify different species using a guidebook/photograph
Count the number of individuals or the population of/ photosynthesis is related to changes in
each type of species present in quadrat light intensity
Count the total number of all the plants present in
quadrat Vary light intensity by altering the distance between lamp
Calculate percentage cover if species of concern is and the plant (seaweed) container
grass.sta Use different strengths of neutral density filters
Use quadrat of the same size OR change the number of lamps with same power ratings
Use same size plot in each area State values of distances in the range 10 to 200 cm
Use the same number of different quadrats/samples per Use sodium hydrogen carbonate for carbon dioxide
plot supply
Replicate the procedure with a different plot in each area Record changes in colour/pH (using pH probe) after a set
Carry out sampling at different times of year/seasons amount of time
Safety: injury/getting lost and staying with a group; allergy Use a control (dead plant/seaweed)
to plants and wearing gloves/protective clothing; allergy OR cover tubes with light proof foil to act as control
to pollen/hay fever and wearing mask or taking Use same volume of hydrogen carbonate solution

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CAIE A2 LEVEL BIOLOGY (9700)

Use same mass of plant/seaweed

2.16. To investigate if there are more
Make a minimum of two replicates and calculate mean
Safety: sodium hydrogen carbonate indicator solution is pollen grains in the atmosphere during
harmful/irritant and wear gloves
the day or night
2.13. To investigate how noise affects
reaction time of human
Carry out experiment in silence (soundproof room)
Ensure consistent/known volume/decibels of noise
Carry out experiment on a large number of volunteers
Do not warn when an action is needed to be taken (e.g. Expose slide/apparatus for a known period of time in
catching an object which is dropped suddenly/needed to different light
make a click on computer keyboard) Use intensities ranging from dark (zero intensity) to light
Use same object/weight of object and dark conditions
Volunteers should be of same or similar age/ same sex Count pollen in the field of view
Use same/dominant hand to catch/make clicks Count at least 3 areas of the slide
Make sure subjects do not take any (named) Measure the diameter of field of view using the eyepiece
drugs/medications/stimulants/depressants graticule
Do not recruit people with conditions affecting reaction
Calculate area of field of view using formula πr2
time (e.g. poor hearing/poor sight/neurological disorders)
Take all readings from the same location
for experiments
Remove any pollen on opening between each slide
Carry out experiments on same time of day
Carry out experiment in an outside location
Make a minimum of three repeats and calculate mean
Make sure there are no walls/hedges/trees in the way
low/medium risk
Repeat the whole investigation on 3 different days and
taking mean
2.14. To investigate if lower epidermis of safety: use mask in case of pollen allergy
leaves has more stomata than upper Low risk investigation

epidermis
2.17. To investigate the optimum
Use strips from upper and lower epidermis temperature for respiration of yeast
Use strips from 5 different leaves of same type of plant
Use microscope and eyepiece graticule Use water-baths at different temperatures (at least 5
Count the number of stomata visible (e.g. in field of view) different temperatures)
Mount epidermis in water/glycerol/(suitable) stain Carry out retesting within the approximate optimum zone
Measure the diameter of field of view using graticule Use methylene blue as indicator
Calculate the area field of view using formula πr2 Keep volume of methylene blue constant
Use graduated pipette to measure volume of methylene
Convert area measured to mm2
blue
The fastest time until blue disappears is optimum
2.15. To investigate how electrophoresis temperature
can be used to obtain genetic Use a control without methylene/same volume of a
solution
fingerprinting Use a standard volume of yeast/suspension in tube
Stir to mix indicator and yeast
Keep samples in wells of agarose gel/support medium Repeats at least 3 times and find mean value/remove
Place wells near the cathode anomalies
Add buffer solution to keep pH constant safety: Toxicity of methylene blue/allergic to yeast
Apply potential difference to buffer Low risk experiment
DNA fragments that move to positive
electrode/anode/DNA is negatively charged
Fragments of different sizes move different distances 2.18. To investigate a method to extract
Smaller fragments move further in given time/faster photosynthetic pigments from algae
and obtain chromatograms

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Use a sample from each type of alga Safety: toxic/irritant/corrosive solvent or dye; wear gloves
Use same number and mass of algae and goggles
Crush algae with a solvent, e.g. ethanol
Find/measure the position of the pigments/colours on the 2.21. To investigate the Km value of
chromatogram and calculate Rf value
Extract pigments from the algae by grinding/crushing enzymes at different
Filter/centrifuge to remove debris/obtain pigments
Concentrate the extract by evaporating or heating
temperatures1/pH2
Run the sample in a solvent for a set distance
Use a suitable range of at least 5 temperatures (10-70
Dry before using second solvent; before solvent front
reaches the end/pre-marked line degrees Celsius)1
Run in second solvent at 90° to first run Use a thermostatically controlled water bath to incubate
Cover the container to prevent evaporation enzyme and substrate solutions at constant
Repeat to compare chromatograms/to find anomalies temperature(s)2
Safety: solvents (toxic/flammable) gloves and goggles Use same concentration of enzyme each time
Use same volume of enzyme each time
2.19. To investigate the activity of free Measure volume using graduated pipette
Use same volume of buffer to maintain a constant pH1
enzymes and immobilized enzymes Add acid/alkali to change pH and measure pH using a

Use same volume/stated volume of enzyme for making probe2

beads and free enzyme incubating enzyme and substrate concentrations
Use methylene blue as indicator separately
Mix enzyme and substrate on the magnetic stirrer and
Use same volume/concentration of methylene blue
Use substrate of same type/volume/surface area immerse a conductivity probe
Measure time using a stopwatch to measure time taken Take readings from meter at same time for each
for colour change temperature
Use thermostatically controlled water bath to keep Carry out experiments at different concentration of
enzymes for the same range of temperature/pH to gather
temperature constant
Use a buffer to maintain pH more data
Allow temperature to equilibrate before mixing enzyme Take a minimum of 3 replicates and find mean
and substrate Safety: enzyme is allergen or irritant; wear gloves and
goggles
Repeat at least 3 times and find mean/identify anomalies
safety: suitable hazard and precaution/low risk
experiment 2.22. To investigate the rate of
transpiration at different light
2.20. To investigate the rate of
intensities using a potometer
movements of pigments with varying
masses in a chromatogram
Count the number of spots
Run the pigment to the same distance moved by solvent
front
Apply the same number of pigments in the origin
Use a capillary tube to give a spot on the
chromatography paper
Draw a base line using a pencil
Concentrate the extract either by drying between
adding spots or evaporating the extract before using
Place the solvent so that the level of solvent is below the
origin line
Cover to prevent evaporation
Dry before spraying with dye
Run at least 3 chromatograms for both enzymes
Calculate Rf value for each experiment Cut/insert stem of plant under water
Take mean of distances travelled by each spot or taking Use petroleum jelly/silicone to make joints air-tight
mean Rf values Remove tube from water to introduce an air bubble

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CAIE A2 LEVEL BIOLOGY (9700)

Obtain a minimum of 5 different light intensities; keep Use same/known/stated volume and concentration of
lamp at a fixed distance and use filters of different nutrient solution/glucose
strengths/thickness Use thermostatically controlled water bath to keep
Allow apparatus/plant to equilibrate before starting temperature constant
measurements Use at least 5 different temperature values between (15
Reset air bubble to start position between measurements degrees Celsius – 80 degrees Celsius)
Measure distance moved by bubble over a set time using Allow temperature equilibration of glucose and yeast
the graduated pipette OR Measure time taken for bubble suspension
to move a set distance using a stopwatch Add a known indicator
Take a minimum of 2 repeats at each light intensity and Measure time taken for permanent colour change
calculate mean/identify anomalies Use a computer-controlled gas flow train to maintain
Low/medium risk investigation: Cut stem and cut away oxygen concentration
from your hand with scalpel Use at least 3 replicates and find mean
Keep temperature constant by carrying out experiment in Identify and eliminate anomalies
a temperature-controlled room Low risk experiment
Carry out experiment in a dark room, no external light Safety: yeast allergenic/indicator irritant, wear mask,
source should be present gloves, and goggles
Use same lamp to ensure same
wavelength/brightness/colour of light 2.25. To calculate respiratory quotient
Keep airflow constant by turning fan off or on at the same
speed by conducting an experiment
Use same species if plant’s leaves/shoot
Use a humidifier to keep humidity the same

2.23. To investigate the effect of

temperature on carbon dioxide
production during respiration of
photosynthetic protoctist
Set up at least 5 water-baths at different temperatures
(state temperature values with units)
Use same volume/concentration of hydrogen
carbonate/indicator to each test-tube
Add oxygen into indicator solution
Ensure same colour/pH of indicator at each test tube
before starting experiment.
Use colorimeter/pH probe to judge colour/measure pH
Use same volume and species of photosynthetic
protoctist
Put indicator and the photosynthetic protoctist in
separate tubes in a water-bath to reach desired Pour known volumes of potassium hydroxide to each
temperature respirometer vessel
Mix protoctist and indicator until endpoint is reached. Add filter paper to act as wicks
Keep pH value/colour of endpoint constant Add known volume of seeds in one of the vessels, make
Use boiled protoctist of the same volume to act as control sure the seeds/invertebrates do not meet potassium
Make at least three replicates and finding mean (of time hydroxide, keep seeds in a box
or pH) and identify anomalies Add same volume of water in the other vessel
Low risk experiment Fit bungs in both vessels
Attach two connecting tubes to vessel with water, one
with a clip on a flexible hose
2.24. To investigate the respiration rates Attach a syringe of known volume and a connecting tube
of yeast of different varieties to vessel with seeds/invertebrates
Add dye/coloured liquid to U-shaped manometer
Use same/known/stated volume of each yeast Connect manometer to the connecting tubes
Add them to separate flasks Make sure that the apparatus is airtight

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CAIE A2 LEVEL BIOLOGY (9700)

Use the syringe (by pressurizing and releasing pressure)

3.1. Measuring sections of specimen
to adjust the manometer so that the fluid levels is same
on both side (e.g. tubules/lumen) with eyepiece
Record position of syringe, position of both sides of
graticule
meniscus and the time
Record new positions of manometer fluid at regular
Set magnification at x40 – x400
intervals. When it reaches the end of one side of the tube, Use stage micrometre and eyepiece graticule
restore original position and record readings on syringe Calculate the number of eyepiece graticule units per
Find volume of oxygen absorbed by seeds/invertebrates
stage micrometre unit/calibrate the eyepiece graticule
in a known period. This is Vol1
with the stage micrometre
Remove potassium hydroxide from both vessels and Convert from eyepiece units/mm to micrometre
wash them out with water Use the formula:
Replace seeds/invertebrates with equal volume of water
and record changes in gas volume for known period. This number of stage micrometer divisions
× value of 1 micr
is Vol2 number of eyepiece graticule units
​

Calculate volume of CO2 produced

Calculate respiratory quotient using formula 3.2. Counting cells using a
Volume of carbon dioxide producedhemocytometer
Respiration Quotent =
Volume of oxygen absorbed
​

Dilute the sample and use a coverslip

V ol1 + V ol2 Count squares where cells are fully inside and touching
RQ = ​

V ol1 the lines.

Calculate the volume of the sample
3. Measurements and Count cells in a sample of known volume
Use microscopes and suitable magnification
counting Calculate the actual number of cells
(Divide the number of cells by the volume) — optional

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