AQA A-Level Biology Required Practical Methods & Notes: Name: Class: Biology Teacher
AQA A-Level Biology Required Practical Methods & Notes: Name: Class: Biology Teacher
Name:
Class:
Biology Teacher:
A-level Biology required practical No. 1
Student Sheet
Casein is a protein found in milk. Trypsin is an enzyme that digests casein. When trypsin is added
to a dilute solution of milk powder, the casein is digested and the solution goes clear.
Method
You are required to find the rate of reaction at five different temperatures. Your teacher will tell
you whether you are going to investigate all the temperatures yourself or whether you will get some
results from other students in your class.
You should read these instructions carefully before you start work.
1. Using a marker pen write an ‘X’ on the glass halfway down one side of each of three test tubes.
2. Add 10 cm3 of the solution of milk powder to each of these three test tubes.
3. Add 2 cm3 of trypsin solution to 2 cm3 of pH 7 buffer in another set of three test tubes.
4. Stand the three test tubes containing the solution of milk powder and the three test tubes
containing trypsin and buffer in a water bath at 20 oC.
5. Leave all six tubes in the water bath for 10 minutes.
6. Add the trypsin and buffer solution from one test tube to the solution of milk powder in another
test tube and mix thoroughly.
7. Put the test tube back into the water bath.
8. Repeat steps 6 and 7 using the other test tubes you set up.
9. Time how long it takes for the milk to go clear. Do this by measuring the time taken to first see
the ‘X’ through the solution.
10. Record the time for each of the three experiments.
11. Using the same method, find out how long it takes the trypsin to digest the protein in the
solution of milk powder at 30 oC, 40 oC, 50 oC, 60 oC.
12. Record your data in a suitable table.
13. Process your data and draw a graph of your processed data.
Notes on this practical:
Notes on this practical:
A-level Biology required practical No. 2
Student Sheet
Preparation of stained squashes of cells from plant root tips; set up and use of and optical
microscope to identify the stages of mitosis in these stained squashes and calculation of a
mitotic index
100 ml beaker
hydrochloric acid (5 mol dm-3)
microscope slide and cover slip
toluidene blue stain
filter paper
mounted needle
scalpel
distilled water
watch glass
forceps
root tip of onion or garlic
microscope and light source.
You are required to prepare a microscope slide of the meristem tissue from an onion root. You will
add a stain to the material which allows you to see the chromosomes. You will look at the slide
under the microscope to identify any cells showing stages of mitosis. You will then calculate the
mitotic index.
Safety
The beaker must be stood on a bench mat. Do not carry the beaker with
acid in it.
N.B. Do not leave root tips for investigation lying about on the bench top prior to staining. Cut
your root tip immediately before you put it into the acid. This will stop any reactions and hopefully
some cells will be in a stage of division.
You should read these instructions carefully before you start work.
1. Stand the beaker on a bench mat before adding approx. 10ml of hydrochloric acid (5 mol dm-3)
2. Place about 2 cm of root tip in the acid and leave for 15 minutes.
3. Set up your microscope while you are waiting.
4. Rinse the root tip in distilled water in the watch glass.
5. Cut off the root tip (1mm) and place on a microscope slide.
6. Cover the section with toluidene blue stain and macerate with the mounted needle to separate
the cells.
7. Continue to macerate until the tissue is well broken and the cells are stained dark blue.
8. Add a cover slip and with gentle finger pressure ‘spread’ the material and blot at the same time
by using a folded filter paper between finger and slide.
9. Look carefully at all slides for cells undergoing mitosis. Chromosomes should stain dark blue.
Repeat for several fields of view.
10. Record your data in a suitable table.
11. Calculate the mitotic index.
Student Sheet
You should read these instructions carefully before you start work.
1. Label six boiling tubes 0, 0.2, 0.4, 0.6, 0.8 and 1.0 mol dm-3 sucrose.
2. Use the 1.0 mol dm-3 sucrose solution and water to make up 20 cm3 of sucrose solution of each
of the following concentrations:
Complete Table 1 to show the volumes of 1.0 mol dm-3 sucrose solution and water that you used to
make up each concentration.
3. Stand the boiling tubes containing the sucrose solutions in a water bath set at 30 oC. Use a
thermometer to check the temperatures in all tubes reaches 30 oC.
4. Using the chipper, cut six chips from your potato tuber. Make sure you remove any peel on the
potatoes. Use a ruler, scalpel and tile to cut all of the chips to the same length. Blot the potato
chips dry with a paper towel, i.e. roll each chip until it no longer wets the paper towel and dab
each end until dry. Do not squeeze the chips. Put each potato chip onto a clean square of
paper towel which you have numbered in the same way as the boiling tubes.
5. Weigh each potato chip. Record these initial masses in a suitable table.
6. At the water bath, set the stop clock to zero. Quickly transfer each potato chip from its square
of paper towel to its own boiling tube with the same number.
7. After precisely 20 minutes, remove the chips from the boiling tubes. Blot the chips dry, as
before. Then reweigh them. Record these final masses in your table.
8. Calculate the change in mass and then calculate the percentage change in mass.
9. Plot a graph of your processed data and use this to determine the concentration of sucrose
which is which has the same water potential as the potato tuber cells.
Table 1
Concentration of
sucrose solution/mol 0 0.2 0.4 0.6 0.8 1.0
dm-3
Volume of 1.0 mol
dm-3 sucrose solution 0 20
/ cm3
Volume of water/cm3
20 0
Notes on this practical:
Notes on this practical:
A-level Biology required practical No. 4
Student Sheet
The effect of alcohol concentration on the leakage of pigment from beetroot cells
Introduction
Beetroot contains high concentrations of betalin. This is a purple pigment found inside the vacuoles
of the cells. The pigment cannot move across undamaged plasma membranes. You will investigate
the effect of alcohol concentration on the amount of pigment leaking through beetroot plasma
membranes.
In Part 1 of the investigation you will produce a set of standards. In Part 2 you will use these
standards to compare the colour of the solutions obtained when beetroot discs have been soaked
in different concentrations of alcohol.
Method
You should read these instructions carefully before you start work.
1. Use the extract and water to prepare a series of six test tubes containing 5 cm3 of different
concentrations of extract. The concentrations should be equally spaced and cover a range from
pure water (0%) to pure extract (100%). These will be your colour standards.
2. Label these standards 0, 2, 4, 6, 8, 10.
3. Complete Table 1 to show the concentration of extract in each tube.
4. Complete Table 1 to show how you made the colour standards in Part 1 of the investigation.
Table 1
Volume of
beetroot
extract/cm3
Volume of water
/cm3
Concentration 0 100
of extract/%
Absorbance
reading from
colorimeter
Student Sheet
Dissection of animal or plant gas exchange or mass transport system or of organ within
such a system
Heart Dissection
a sheep’s heart
dissecting tray and board
dissecting instruments
labels and pins.
You should read these instructions carefully before you start work.
2. Cut down each side of the heart to open up the left atrium and left ventricle and the right atrium
and right ventricle.
Look for the tendinous cords holding the atrio-ventricular valves, and lift the weight of the heart
by holding one of these cords over a dissecting needle.
Look how thin the atrio-ventricular valves are.
Examine the thickness of the walls of the ventricles.
Which side is thicker, and why?
Look at the walls of the atria, they are much thinner, can you think why?
Push the handle of the dissecting needle up behind the atrio-ventricular valves. You should
notice that the aorta and pulmonary arteries cross over.
3. Make some little flags from pins and sticky labels and label the parts of the heart that you can
identify. Make sure they are legible and visible as you look down on your dissection.
Ask your tutor to check your labeling and take a photograph so you can include it in your notes.
Packing away:
Use the disinfectant spray to clean the dissecting board and bench, using paper towels to dry
them. Dispose of the towels in the yellow disposal bag along with your plastic gloves.
Notes on this practical:
A-level Biology required practical No. 6
Student Sheet
Aseptic technique producing bacterial plates and use of mast ring of antibiotics
Method
You should read these instructions carefully before you start work.
1. Spray the bench with the disinfectant and wipe down with paper towels. Place the sterile plastic
sheet on the cleaned bench.
2. Place the Bunsen burner on a heat proof mat and light it.
3. Place the agar plate, the McCartney bottle and the spreader next to the Bunsen burner.
4. Write your name, the date and the name of the bacteria on the underside of the agar plate.
5. Wash your hands.
6. Remove a sterile 1 cm3 pipette from the foil and place the filler onto it.
7. Flame the neck of the McCartney bottle.
8. Dip the pipette into the bottle and remove 0.3 cm3 of the bacterial culture.
9. Flame the neck of the bottle again and replace the lid.
10. Lift the lid of the agar plate at an angle facing the Bunsen burner with your left hand. With
your right hand, squeeze the contents of the pipette onto the surface of the agar.
11. Replace the lid of the agar plate and place the pipette into the beaker of disinfectant.
12. Take the sterile plastic spreader in your right hand. Facing the Bunsen, lift the lid of the agar
plate and use the spreader to make sure that the bacteria are evenly spread around the
surface of the agar.
13. Replace the lid of the plate place the spreader into the beaker of disinfectant.
The disks you will be using have eight arms, each arm containing a different anti-bacterial agent.
These are coded as follows:
1. Take a pair of forceps. Only handle the Multodisk with the forceps.
2. Remove a Multodisk from its tin and transfer it to the centre of the agar plate.
Do not hold the disk by one of its arms.
3. Carefully flatten the Multodisk onto the surface of the plate, using the forceps.
Place the forceps into the beaker of disinfectant.
4. Hold the lid of the plate in place with two pieces of tape.
5. Place your plate upside down in an incubator at 25oC for 48 hours.
6. Now wash your hands.
After incubationCaution - plates must not be opened after they have been incubated
7. Examine your plate and try to identify the colonies which have not been able to grow near the
Multodisk arm(s). These are called zone(s) of inhibition. Turning the plate upside down and using
a ruler measure the diameter of the zones of inhibition. Calculate the area of the zone of inhibition
using the formula
Student Sheet
Use of chromatography to investigate the pigments isolated from leaves of different plants
eg leaves from shade-tolerant and shade intolerant plants or leaves of different colours
Introduction
In plants, chlorophyll is the main pigment that absorbs light energy during photosynthesis. Most
plants have other photosynthetic pigments as well and these are not green. You will be using a
technique called chromatography to separate chlorophyll and other pigments from two different
leaves, A and B.
Method
boiling-tube rack
two boiling tubes with bungs
small glass measuring cylinder
solvent
chromatography paper
glass rod
two leaves, A and B
cork borer
tile on which to use cork borer
ruler
pencil
drawing pins
marker pen
sticky tape.
Safety
You should read these instructions carefully before you start work.
1. Set up two boiling tubes at the start of the investigation. Add 3 cm3 of solvent to each of the
two boiling tubes. Put a bung in the top of each tube and stand them upright in a rack. Label
the tubes A and B.
2. Take a piece of chromatography paper that fits into the boiling tube, as shown in the diagram.
Rule a pencil line 2 cm from the bottom of the filter paper. This line is called the origin. Write
leaf A at the top of the chromatography paper in pencil.
3. Cut a disc from leaf A with a cork borer. Try to avoid the veins and midrib of the leaf when you
do this.
4. Place the leaf disc on the chromatography paper at the centre of the line marking the origin.
Crush the disc into the paper with the end of a glass rod. The crushed leaf disc should leave a
stain on the chromatography paper.
5. Pin the chromatography paper to the bung with a drawing pin, and then put the
chromatography paper into the tube labelled A as shown in Figure 1. Make sure the end of the
chromatography paper is in the solvent and that the solvent does not come above the origin.
Put the tube carefully back into the rack and do not move it again.
Figure 1
6. Let the solvent run up the chromatography paper until it almost reaches the top of the paper.
Remove the chromatography paper from the tube and immediately draw a pencil line to show
how far the solvent moved up the paper. This line marks the solvent front.
7. Replace the bung in the tube.
8. The filter paper with its coloured spots is called a chromatogram. Let the chromatogram dry.
Using a pencil, draw round each coloured spot on the chromatogram.
9. Repeat step 2 with the second piece of paper but write B at the top of the chromatography
paper.
10. Repeat steps 3 – 8 with leaf B.
Calculate the Rf value for each of the pigment spots on each chromatogram.
Student Sheet
Investigation into the effect of a named factor on the rate of dehydrogenase activity in
extracts of chloroplasts
The effect of ammonium hydroxide on the time taken for chloroplasts to decolourise DCPIP
In this investigation you will use a chloroplast suspension and a blue dye called DCPIP to monitor
the rate of dehydrogenase activity. DCPIP goes from blue to colourless when it accepts electrons
released by the chlorophyll.
Method
spinach leaves
access to a blender
measuring cylinder
muslin ( or material for filtering)
filter funnel
3 beakers
ice
isolation medium (cold)
DCPIP solution (cold)
distilled water (cold)
ammonium hydroxide solution (cold)
test tubes
test tube rack
syringes (1cm3 and 5 cm3 )
piece of aluminium foil
lamp
marker pen
timer.
You should read these instructions carefully before you start work.
Student Sheet
Investigation into the effect of a named variable on the rate of respiration of cultures of
Yeast is a single- celled fungus. It can respire aerobically and anaerobically. During aerobic
respiration, the transport of electrons is linked to the synthesis of ATP. In this investigation these
electrons will be accepted by a substance called methylene blue. When methylene blue accepts
electrons, it changes from blue to colourless.
Method
You should read these instructions carefully before you start your investigation.
Student Sheet
Method
Control experiment
If the left and right halves have no effect on the distribution of the maggots the expected results
would be six in each half, but this will not always occur because of chance distribution. If your
results are not 6 in each half do a statistical test on your results to discover the probability of them
occurring by chance. If this test shows a greater than 5% probability of the results occurring by
chance then you can proceed with the experiment.
1. Cover half the choice chamber with black paper to make it dark.
2. Place 12 maggots in the chamber through the central hole, using the teaspoon.
3. Wait 4 minutes and then record the number of maggots in the dark and the light halves.
If light has no effect on the distribution of maggots the expected results would be six in each half.
Now do a statistical test on your results to find the probability of them occurring by chance.
1. Place damp paper towel in one half of the choice chamber and silica gel in the other. Use the
humidity test strips to ensure that a humidity gradient exists in the chamber before adding the
maggots. Use the forceps to place the humidity test strip.
2. Place 12 maggots in the chamber through the central hole.
3. Wait 4 minutes and then record the number of maggots in the humid and the dry halves.
In reality living organisms do not have simple choices between one pair of contrasting
environmental factors. If you have time do a final experiment with the choice between dark and
dry, dark and humid, light and dry, light and humid. Again test the probability of your results
occurring by chance with a statistical test.
You should read these instructions carefully before you start work.
1. Cut out pieces A, B and C from the card by cutting along all the solid lines.
2. Fold along the dashed and dotted lines, keeping dashes on the inside and dots on the outside.
3. Glue the tabs to form the maze shown in Figure 1.
4. Cut out the barrier (piece D) and place it at the position shown in Figure 2.
Figure 2
This experiment should give equal numbers turning left and right. This section of the maze could
be used to investigate the effect of variables such as light by covering one side of the maze with
black paper and then the other.
Many animals show behaviour called turn alternation. This means if the animal is forced to turn in
one direction it is more likely to turn in the opposite direction next time it has a choice. The maze
can be uses to allow you to investigate whether maggots show turn alternation.
Risk assessment
Risk assessment and risk management are the responsibility of the centre.
Trialling
Additional notes
The Bluebottle adult flies hatching from the maggots are classed as statutory nuisance animals by
DEFRA (listed in document “Nuisance Insects and Climate Change” March 2009). The maggots
should therefore be killed (e.g. by placing in a freezer for a week) before wrapping securely and
placing in a bin collected directly by refuse collectors. Animals that have been used in experimental
work are regarded as animal by-products and as such should not enter the food chain. For
disposal, the flies should be killed as described above, and placed in the normal refuse.
See Hazcard 25 for Cobalt chloride papers, and found in self-indicating silica gel, causes skin
sensitisation. The chemical is also potentially carcinogenic by inhalation. Students should use
forceps to place the cobalt chloride paper in the choice chamber. Students should not directly
handle the papers or self-indicating silica gel.
Notes on this practical:
Notes on this practical:
6.
A-level Biology required practical No. 11
Student Sheet
Method
1. Label the test tubes with the name of the patient and add 2 cm3 urine samples from each
patient.
2. To each test tube add 2 cm3 Benedict’s solution. Mix the contents of the tube.
1. Label six test tubes 0 to 10 mmol dm-3 as shown in the table below.
2. Dilute the glucose standard (10 mmol dm-3) with water in the labelled test tubes and complete
the table to show volumes used to achieve each concentration.
3. Add 2 cm3 of Benedict's solution to each tube. Mix the contents of each tube.
4. Place all the test tubes into the water bath together (including the tubes with the urine samples)
and time for four minutes. Allow to cool before taking readings from the colorimeter.
5. Use the contents of the 0.0 mmol dm-3 glucose solution tube, which you have heated with
Benedict's, as a blank to calibrate the colorimeter to zero absorbance. Place the remaining
samples in cuvettes into the colorimeter and read the absorbance.
6. Record your results in a table and plot a graph of the absorbance of the known concentrations
of glucose.
7. Using the graph and the absorbance values obtained for the urine samples read off from the
graph the concentration of glucose in the urine samples.
8. Record your results in a suitable table.
Notes on this practical:
Notes on this practical:
A-level Biology required practical No. 12
Student Sheet
Investigation into the effect of a named environmental factor on the distribution of a given
species
Investigation into distribution of dandelions in a lawn not treated with herbicide and a lawn
treated with herbicide using a point quadrat
Method
You should read these instructions carefully before you start work.