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- Precise: A measurement with very little spread about the mean value. One with a lot of
decimal places.
- Reliable: The results can be repeated. The reliability of data within a single investigation can be
improved by carrying out repeat measurements.
- Valid: Measurements made are affected by a single independent variable only. They are not
valid if the investigation is flawed and control variables have been allowed to change.
· When drawing a table to record data, be sure to include the original observed measurements
that would be recorded during the experiments. You might include additional columns for
calculated values such as a difference or percentage change.
· Hypotheses and statistical tests make use of specific and precise scientific language. Use the
correct terminology of significant difference (T test – when testing two different discrete data sets)
or significant correlation (Spearman’s Rank correlation test – when testing for correlation between
two variables) to formulate a good null hypothesis.
· Make sure that you directly answer the question. If you are describing changes from the usual
course of events, be specific about the changes and describe their nature by using terms such as
more / fewer / greater / slower.
· When describing a method the phrase 'record the results' is very vague and should be avoided.
Specify exactly what should be recorded: for example, in the case of habituation, record the time
taken for the snail to fully re-emerge from its shell.
- When measuring the colour of the solution make sure to use a suitable reference corvette /
solution
UNIT 1
· Why Daphnia?
- 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
- Transparent so easy to measure heartbeat
· Method:
- Independent variable: Caffeine concentration
- Dependent variable: Heart rate of Daphnia
- Place a Daphnia in each of 5 different solutions (4 different concentrations of caffeine and one
with distilled water to act as a control)
- Leave Daphnia for 5 minutes to acclimatise
- Immobilize the Daphnia using a little cotton wool in a cavity slide and observe under
microscope
- Count and record the no. of heartbeats in one minute
- Repeat 5 times at each concentration and allow means to be calculated
· Variables to be controlled:
- Mass of fruit / age / source / …
- Time for storage
- Method of juice extraction
- Volume / concentration of juice / DCPIP
- Temperature
- Same end point colour
· Method:
- Independent variable: Fruit juice
- Dependent variable: Volume of juice required to de-colourise 1cm3 of DCPIP
- Put 1cm3 of DCPIP solution into a test tube
- 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
- Repeat 5 times for each juice to calculate means
- 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.
- Concentration 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
· Limitations:
- Difficulty in controlling temperature
- End point difficult to judge as needs to be just when blue colour disappears especially in highly
coloured juices
- Some loss of solution when transferring from one beaker to another
- Accuracy of measuring equipment
· Factors that affect the permeability of the beetroot cell membrane are:
- Temperature
- Age
- Storage
- Duration
- Prior treatment with solvent
- pH
- Bile salts
· Method:
- Independent variable: Temperature of water
- Dependent variable: % transmission of light through resulting solution
- 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)
- Leave for 15 minutes
- Remove beetroot and shake tubes to disperse dye.
- Calibrate the colorimeter using distilled water in a cuvette as a reference / control.
- Take readings of absorbance of the water in the tubes
- Take 5 repeats at each temperature and calculate means
· Results and reasons: As temperature increases, % transmission slightly increases. This is due to
membrane molecules gaining more heat energy and vibrating more, creating large gaps in the
membrane that enable dye to be released. Proteins in membrane may become denatured, leaving
large pores through which the dye leaks
· Limitations:
- Pigment is not equally distributed throughout the beetroot
- Some beetroot may have skin on affecting surface area
- Size of beetroot is difficult to control
· 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
- Repeat at least 5 times at each concentration and calculate means
UNIT 2
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
- Break up the tips gently using a mounted needle on a microscope slide
- Add toluidine blue to stain the chromosomes, warming if needed to intensify the stain
- Place a glass coverslip on top and squash gently
- Observe under the microscope
· Variables to be controlled:
- Same age / size seeds
- All seeds should come from the same plant
- Temperature
- Light Intensity
- Time allowed to grow
- Atmospheric carbon dioxide concentration
- Humidity
· 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
· Variables to be controlled:
- Length of fibre
- Size of each individual mass
- Width / diameter / cross-sectional area of fibre
- Fibres must come from the same plant species
- Same age
- Same parent plant to reduce genetic variation
- Same prior treatment
- Temperature
- 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
- Record the mass at which the fibre breaks
- Repeat 5 times at each thickness to calculate a mean
· Preliminary Work:
- See if proposed method will work
- See if the plant chosen will grow in hydroponic unit
- Select a range of concentrations
- Check for suitable conditions for digestion e.g. temperature, pH
- Find a suitable method of measuring growth
- Check for most suitable conditions for growth of plants
- Select suitable time scale for measuring growth / stain digestion
· Things affecting enzyme action are: protein type, volume of solution, stirring, pH, temperature,
surface area, protein concentration…
· Variables to be controlled:
- Volume of mineral solution
- Concentration of solution
- Species of plant
- Light intensity
- Age / size of plant / seedling
- Genetically similar\same parent plant
- Temperature
· 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
- Independent variable: Concentration of magnesium. Range of suitable concentrations
suggested (at least 5) (0 (control), 10%, 30%, 50%, 70%, 90%, 100%)
- 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
- Leave the tubes for a week
- Record changes in mass / height / root length
- 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
· 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
· 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 cellotape 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:
- Practise proposed method / see if proposed method will work
- Allows selection of appropriate species of frog to be worked out
- Carry out experiments to determine a suitable method for collecting secretions from frog
- Carry out experiments to determine appropriate concentration / volume of frog secretion
- 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 / …
- Determine best method of measuring dependent variable
· Variables to be controlled:
- Concentration of plant material / antibiotic
- Lawn of bacteria on petri dish
- Same volume of plant material / antibiotic on each disc
- Disc size
- Bacterial species (E. Coli)
- Temperature
· Method:
- Dependent variable: Zone of inhibition / absorbance of culture
- Independent variable: secretions from different frogs / antimicrobial solution
- 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
- Dip small disk of blotting paper and allow to dry
- 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
- Incubate at 25 / 30 ºC for a week
- 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
- Repeats taken and means calculated
· Limitations:
- Difficult to control all variables (affecting bacterial growth)
- Other components of secretions may affect bacterial growth masking the effect of the
antibiotics
- Difficult to standardise extraction of secretion
- Other variables related to frog e.g. age, size, gender
- Uneven spread of bacteria
- A variable may be acting as a limiting factor for bacterial growth (give example)
- Need to test effect on more than one type of bacteria
UNIT 4
· 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
2. Count number of chosen species in each quadrat or estimate % abundance
3. Density = Total no. of plants counted / (Area of one quadrat x Total no. of quadrats taken)
- 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:
- Check for other variables that need to be taken into account / controlled
- Light intensity
- O2 concentration
- Mineral Concentration
- Water
- Food
- Time
- pH
· Method:
- Dependent variable: E.g. percentage change in mass of plant tissue / no. of hatched shrimp /
height for seedlings
- 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
· 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 attach 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 synthesises 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 extension / 70 to 80 ºC / step 3 à
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
· Collect and analyse 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
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)
UNIT 5
6. Investigating Respiration
· Method:
- 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
- Time
· 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.
· 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
- The apparatus can be calibrated so that the movement of the lid corresponds to a given
volume.
- 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.)
· 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.
· Ethical issues:
- 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
· Method:
- 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
· 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
· Calcium ion involvement in habituation: Repeated stimulation affects calcium channels. Fewer
calcium ions enter the pre-synaptic membrane and so less neurotransmitter is released into the
synaptic cleft. Less depolarisation of the post-synaptic membrane will occur and fewer sodium
channels will open. No action potential will be generated and so no impulse sent, no response is
observed.
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