Professional Documents
Culture Documents
- Daphnia are small and show the results of the experiment quickly
- They have simple nervous systems so are less likely to feel pain
- They are abundant so easy to get hold of / No damage to environment when a few are removed
from it
- Temperature
- Volume of solution
- Time of acclimatization
- Counting time
· Method:
- Place a Daphnia in each of 5 different solutions (4 different concentrations of caffeine and one with
distilled water to act as a control)
- Immobilize the Daphnia using a little cotton wool in a cavity slide and observe under microscope
Increasing caffeine concentration causes the electrical activity of the sinoatrial node to increase, making
it depolarize. As it depolarizes, the right and left atria contract and the impulse travels to the
atrioventricular node where, after a delay of about 0.13 seconds, the impulse continues to travel
towards the ventricles. This delay ensures that the atria have finished contracting and ventricles are full.
The signal then reaches the Purkyne fibres that conduct the impulses to the apex of the ventricles where
contraction begins and travels upwards towards the atria.
Caffeine also affects the ventricles, leading to an increase in the rate of contraction and relaxation of
each heartbeat. This means that, as well as beating faster, the heart's individual beats are associated
with an increased cardiac output.
· Variables to be controlled:
- Temperature
- Fill a plastic syringe with juice and add drops to the DCPIP until the blue of the DCPIP is lost. Record
the volume of juice added
- To calculate the actual Vitamin C concentration, the DCPIP solution must be calibrated. A solution
of known Vitamin C concentration is added to 1cm3 of DCPIP until it is decolourised and the volume
recorded.
- Conc. of Vitamin C in juice = (Con. of Vitamin C solution x Volume of Vitamin C solution needed to
decolourise 1cm3 DCPIP) ÷ Volume of fruit juice needed to decolourise 1cm3 DCPIP
Edexcel IAL Unit 6
· Limitations:
- End point difficult to judge as needs to be just when blue colour disappears especially in highly
coloured juices
· Factors that affect the permeability of the beetroot cell membrane are:
- Temperature
- Age
- Storage
- Duration
- Prior treatment with solvent
- pH
- Bile salts
· Variables to be controlled are:
- pH
Edexcel IAL Unit 6
· Method:
- Using a cork borer and knife, cut 5 pieces of beetroot equal in mass and size
- Rinse the beetroot pieces with water and gently pat them dry with tissue before using them as,
when cutting, pigment is released from broken cells and must be removed before starting or solutions
will be darker than they should
- Place one piece into each of 5 tubes and add 5 1cm3 water to each one
- Place each tube into a water bath of different temperature (e.g. 15, 20, 25, 30, 35)
· Limitations:
- Volume of enzyme
- Volume of substrate
- Concentration of substrate
- pH
· Method:
- Take 5 test tubes. In 4, place increasing volumes of trypsin solution e.g. (1, 2, 3, 4 cm3 ) and make up
the volume to 4 cm3 using distilled water. The other test tube should be filled with 4 cm3 of water to act
as a control.
- Add 5 of milk powder (casein solution) as substrate and start the stopwatch
- Measure the cloudiness of the solution over time using a colorimeter (every 30 secs for 10 minutes)
against water as a reference / control
· As concentration of enzyme increases, rate of reaction increases up to a point, where all enzyme has
metabolised all substrate immediately
At low temperatures the reaction is slow because the enzyme and substrate molecules don’t collide very
often and move slowly. Increasing temperature increases kinetic energy and so frequency of collisions.
The substrate binds to the enzyme’s active site more often thus increasing the rate of reaction. After the
optimum temperature bonds holding the 3D – shape of the enzyme together start breaking so it loses its
shape and the enzyme-substrate complex can no longer form. The enzyme is denatured.
Edexcel IAL Unit 6
5. Observing Mitosis
· Safety:
- Risk of injury to hands by sharp knife or mounted needle so wear thick gloves or cut away from
body
- Acid is corrosive so wear gloves to reduce risk of injury and safety glasses to reduce risk of injury to
eyes
- Stain may stain clothes and skin so wear gloves and lab coat
- Glass coverslip may break and cut your fingers so wear gloves to protect hands
· Method:
- Cut the last 0.5 cm off the end of actively growing garlic or onion root tips
- Treat with acid to soften tissue by breaking down the middle lamella so that the cells will separate
easily when squashed
- Add toluidine blue to stain the chromosomes, warming if needed to intensify the stain
· Method:
- Use week-old mustard seedlings. Cut off the top 2cm (stem and leaves) and suspend in agar in a
test tube
- Leave for a week and look for new roots / leaves forming
- Cells at the bottom of the stem differentiate to become new roots which demonstrates
pluripotency
· Safety:
- The fibre could enter and injure the eye when it snaps so wear safety glasses to protect eyes
- Place layers of cloth beneath the mass hanger to stop masses from falling onto the foot and injuring it
Edexcel IAL Unit 6
· Variables to be controlled:
- Length of fibre
- Humidity
· Method:
- Soak nettle plant stems in water for a week to soften the tissues and allow the fibres to be easily
extracted
- Select adequate fibre (taking into account all variables) and attach one end to a clamp and stand
then progressively hang masses on the other end
· Safety:
· Preliminary Work:
· Things affecting enzyme action are: protein type, volume of solution, stirring, pH, temperature,
surface area, protein concentration…
· Variables to be controlled:
- Concentration of solution
- Species of plant
- Light intensity
- Temperature
Edexcel IAL Unit 6
· Method:
- Dependent variable: E.g. mass of plant tissue, mass of fruit, length of shoot, number / colour of
leaves. Description of method of measuring change in dependent variable
- Take six plants / seedlings and place each of them into a test tube with a different concentration of
solution (one with distilled water to act as a control)
- Cover each tube with foil to exclude light and prevent algae growth that could affect concentration
of mineral ions
- Repeat at each concentration / for each mineral ion 5 times and calculate mean
- Use of graph to identify other values of concentration to test to identify optimum concentration
- Use of graph to estimate range for optimum / to identify other values of concentration to test to
identify optimum concentration
- Concentrations above those first reaching maximum rate of digestion would be wasteful
· Limitations:
- Difficult to control all variables affecting plant growth / protein digestion e.g. seeds do not
germinate at the same time, genetic differences between the plants… / surface area of stain, protein
concentration
- Limiting factor(s)
- Experimental conditions may not match those normally used
- More than one type of mineral for effective growth of plants
- Difficult to measure the dependent variable
Edexcel IAL Unit 6
· Ethical Issues:
- Welfare of frogs e.g. frogs should be kept in suitable conditions / not be harmed when collecting
secretions
- Avoid skin contact with frogs e.g. wear gloves when handling them / wash hands after handling
them / wear eye protection
· Safety:
- Wipe working area with antiseptic solution / work close to a Bunsen Burner which sets up
convection currents of sterile air to prevent growth of unwanted harmful bacteria / contamination
- Secure lids with cello tape but don’t seal completely in order to avoid pathogenic anaerobic
bacteria to grow
- Don’t use 37 ºC as this is human body temperature and could encourage pathogenic bacteria to grow
· Preliminary Work:
- Carry out experiments to determine a suitable method for collecting secretions from frog
- Carry out experiments to determine the most appropriate method of applying the secretions to the
plates
- Carry out experiments to determine the best parameters for another named variable e.g. suitable
timescale for measuring the inhibition of bacterial growth / conditions for growth of the bacteria / type
of bacteria / …
· Variables to be controlled:
- Disc size
- Temperature
· Method:
- Prepare, under sterile conditions, petri dishes with a thin layer of agar in them
- Once the agar is set, spread a drop of bacterial culture (E. Coli) over the surface using a sterile glass
spreader to form a lawn
- Prepare the extract by crushing material using a pestle and mortar with alcohol if necessary
- Minimally lifting the lid, place on the centre of the agar and press lightly
- Secure lids with 2 pieces of cellotape but don’t seal completely in order to avoid pathogenic
anaerobic bacteria to grow
- Observe the plates and the zone of inhibition will be clear. Measure its diameter to give an idea of
relative antimicrobial strength / effectiveness against microbes
· Limitations:
- Other components of secretions may affect bacterial growth masking the effect of the antibiotics
- A variable may be acting as a limiting factor for bacterial growth (give example)
· Safety:
· Ethical:
· Preliminary Work:
· Sampling Methods:
- Random Sampling: Used when measuring density of a plant species or slow moving animals.
1. Set up grid using tape measure and use random numbers to generate points to place at least 10
quadrats
3. Density = Total no. of plants counted / (Area of one quadrat x Total no. of quadrats taken)
Edexcel IAL Unit 6
- Systematic Sampling: A line transect is used to study changes in plant species across an area.
3. Record data
- Light intensity
- Surrounding vegetation
- Slope
- Temperature
- Soil water
- Humidity
- O2 concentration
- pH
- Clear table which matches method description with headings and units
· Limitations:
- Difficult to control all variables (abiotic factors affecting the variables being investigated)
- Laboratory conditions may not relate to what happens in reality/real life situation
- We assume that the species is evenly distributed throughout the area and that the placing of the
quadrats is entirely random
- Movement of organisms
· Safety:
· Preliminary Work:
- Practise proposed method to see if proposed method will work
- Determine appropriate dependent variable
- Check most suitable conditions for growth of plant tissue
- Select suitable timescale for measuring growth rates
- Check for other variables that need to be taken into account / controlled
- Check if the type of plant / tissue used is affected
· Method:
- Dependent variable: E.g. percentage change in mass of plant tissue / no. of hatched shrimp / height
for seedlings
- Independent variable: Concentration of plant growth regulator / temperature (at least 5 different
ones)
- Specific descriptions of plant tissue culture provided (e.g. need to grow on nutrient gel, aseptic
conditions, antibiotics in gel to prevent growth of microorganisms…) Same as for Totipotency and Plant
Tissue Culture
· Limitations:
- Difficult to control all variables affecting tissue growth + example e.g. exposure to bacteria
- Need for more than one type of plant growth regulator for effective growth
1. Put an equal number of brine shrimp eggs in water baths set at different temperatures
2. Make sure all other variables are the same for each water bath
3. The number of hatched brine shrimp in each water bath are recorded every five hours- the hatch rate
in each water bath can be calculated using: number of hatched brine shrimp in each water bath/number
of hours
· Method:
- A mixture is prepared containing: the DNA sample, DNA polymerases (with v. high optimal
temperatures), DNA primers (short, single-stranded lengths of DNA that are complementary to those at
the start of the STRs, they have fluorescent markers attached that aid the production of the final profile)
and nucleotides
- The mixture is the placed into a PCR machine where it undergoes the following cycle
1. Sample is heated to 95 ºC à This separates the double helix into two strands
2. Mixture is cooled to 55 ºC à Allows the primers to bind to the start of the STRs
3. Further heating to 70 ºC à DNA polymerases attaches to the primers and extend them, replicating
the STR sequence and the adjacent DNA
4. Cycle is repeated for about 25-30 times, which takes about 3 hours, to produce a mixture of
different-length fragments unique to the individual
· The properties of the enzyme relevant to its biological activity in the amplification
process:
- It synthesizes a new strand of DNA complementary to the template strand in one direction. A
primer is needed to begin synthesis of the complementary strand.
- In the context of denaturation of DNA / 90 to 95 ºC / step 1 If temperature too low DNA strands will
not separate.
- In the context of primer annealing / 40 to 70 ºC / step 2 If temperature is too high less binding of
primers will occur
- In the context of extension / 70 to 80 ºC / step 3 If temperature is too low synthesis of new DNA
strands would not be completed à If temperature is higher than 95 ºC, the enzyme will denature
Edexcel IAL Unit 6
· Collect and analyze samples from more than one individual of each species because:
- There will be genetic variation between individuals of the same species, testing more than one
sample will control for these differences
4. Gel Electrophoresis
After using PCR, gel electrophoresis can be used to separate them according to their size and an image
of the fragments produced
· Method:
1. The sample mixture is mixed with a coloured dye and placed carefully into wells at one end of
agarose gel
2. The gel is immersed in a buffer solution in a tank and a potential difference is set up across it
3. DNA is –vely charged so will move towards the +ve electrode at the other end of the gel
4. Smaller fragments will move faster so mixture is separated out into a pattern of bands
5. The wells have a reference mixture of DNA fragments of known length to compare your samples to
- Seeing the fragments:
1. Fluorescent primers glow under UV light and allow a gel photo to be taken
2. DNA can be transferred from the gel to a nylon membrane by Southern blotting, which can be
treated with a DNA probe. This binds to the bands and carries either a fluorescent or radioactive marker.
Radioactive ones can be seen using autoradiography
3. Coloured DNA probes can be added to gels to see the bands directly
- Graphs can be plotted of the size of fragment against level of fluorescence (gives abundance of
fragment)
Edexcel IAL Unit 6
6. Investigating Respiration
· Method:
- Allow time for them to acclimatise to their surroundings and then move the drop of coloured liquid
back to 0 on the scale using a syringe
- Start the stopwatch and note the position of the coloured liquid at regular intervals of 5 minutes.
Subtract the final value from the first to give the overall distance moved.
- Type/source of seeds
- Mass/number of seeds
- Age of seeds
- Ph
- No. of organisms
· Yeast will respire faster using glucose because glucose is the starting point for glycolysis reactions in
respiration; it is the first molecule to be phosphorylated. / Yeast will respire sucrose faster because it
can be broken down into molecules of glucose and fructose; providing double the substrate for
glycolysis / Yeast will respire sucrose more slowly because sucrose needs to be hydrolysed to glucose
and fructose in order to be used in glycolysis. / Rate of uptake of sugars differs: larger molecules may be
taken up more slowly.
Edexcel IAL Unit 6
· Effects of:
- No oxygen during investigation à No/less movement of the liquid in the respirometer. No/less
change in volume/pressure of the gas. Aerobic respiration stops / Anaerobic respiration takes place.
Anaerobic respiration produces no carbon dioxide.
- Increasing temperature à Will increase rate of respiration (as it is enzyme controlled) but also the
volume of air in the apparatus
Using a respirometer
1. Each tube (test and control) contains potassium hydroxide solution which absorbs carbon dioxide
2. The control tube is set up in exactly the same way as the test tube, but without the woodlice, to make
sure the results are only due to the woodlice respiring
3. The syringe is used to set the fluid in the manometer to a known level
4. The apparatus is left for a set period of time (20 mins)
5. During that time there'll be a decrease in the volume of air in the test tube, due to oxygen
consumption by the woodlice
6. The decrease in the volume of the air will reduce the pressure in the tube and cause the coloured
liquid in the manometer to move towards the test tube
7. The distance moved by the liquid in a given time is measured, this value can then be used to calculate
the volume of oxygen taken in by the woodlice per minute
· Method:
This uses a spirometer. Adding air to the chamber makes the lid of the chamber rise in the water, and
removing air makes it fall. Movements of the chamber are recorded using a kymograph (pen writing on
a rotating drum). The volume of air the person inhales and exhales can be calculated from the distance
the lid moves.
- The apparatus can be calibrated so that the movement of the lid corresponds to a given volume.
- The chart recorder can be set to move at a known speed.
- A canister containing soda lime is inserted between the mouthpiece and the floating chamber. This
absorbs the CO2 that the subject exhales.
- A disinfected mouthpiece is attached to the tube, with the tap positioned so that the mouthpiece is
connected to the outside air. The subject to be tested puts a nose clip on (to ensure no breathing occurs
via the nose), places the mouthpiece in their mouth and breathes the outside air until they are
comfortable with breathing through the tube.
- Switch on the recording apparatus and at the end of an exhaled breath turn the tap so that the
mouthpiece is connected to the spirometer chamber. The trace will move down as the person breathes
in. After breathing normally the subject should take as deep a breath as possible and then exhale as
much air as possible before returning to normal breathing.
- Repeats could be for same student at same time each day for a week or with 10 different students
(same age, gender, health, etc.)
Edexcel IAL Unit 6
· Variables to be controlled:
- Temperature
- Standardise exercise
· How breathing is controlled by the nervous system in response to changing positions e.g.
standing up and sitting down: More energy is needed when standing up. The sympathetic nerve
increases heart rate. The ventilation centre in the medulla responds to chemoreceptors in the carotid
that detect changes in levels of carbon dioxide in blood. Motor cortex. Nerve impulses go to muscles
involved in breathing.
1. A spirometer has an oxygen-filled chamber with a movable lid
2. A person breathes through a tube connected to the oxygen chamber
3. As the person breathes in the lid of the chamber moves down, when they breathe out it moves up
4. These movements are recorded by a pen attached to the lid of the chamber, this writes on a rotating
drum, creating a spirometer trace
5. The soda lime in the tube the person breathes into absorbs carbon dioxide
· Ethical issues:
· Safety:
- Snail secretions may irritate skin or cause allergies or carry microbes à hands should be washed
thoroughly before and after handling snails
- Temperature
- Background Noise
- Humidity
- Light Intensity
- Species
- Age
- Gender
Edexcel IAL Unit 6
· Method:
- Dependent variable: Time taken to fully emerge
- Independent variable: Number of pokes
- Place snail on clean, firm surface
- Allow time until snail has fully emerged from shell and has acclimatised
- With a moistened cotton wool bud, firmly but carefully touch the snail between the eye stalks,
starting the stopwatch immediately
- Record the time taken for the snail to fully re-emerge
- Repeat the touch and timing for a total of 10 touches
· Outcome: As the number of stimuli increase, the time taken for the snail to re-emerge decreases.
· Limitations:
- Snails already handled before the experiment may not react in the same way
- Determining when a snail has fully emerged
- Lack of moisture may encourage snail to stay more in its shell
· An increase in temperature increases the rate of chemical reactions in the snail, allowing it to re-
emerge more quickly. To control temperature use incubator/conditioner at constant temperature
suitable for snail