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A2 BIOLOGY CORE PRACTICAL SUMMARY

Name of practicalIndependent &dependentvariablesOther variablesto be


controlledequipment Method and outcome Possible evaluation issues
Observing patternsby Ecological sampling
Random samplingSystemic samplingAbiotic factorse.g. light,temperature, soilwater, humidity,O
2
concentration,pH, aspect, slopeangleGridded QuadratTape measurePoint quadratPitfall trapSweep NetPooterTullgren
funnelBaermann funnelSeveral methods.1
random sampling
= set up grid using tape measure, use random numbers to generate points to place quadrat to collect data.2
systemic sampling
= line transect often used especially to study zonation. A tape measure is laid along several zones to belooked at and
quadrats are used to record data at regular intervals
3 Measuring abundance
Density = presence of organisms per quadratFrequency = percentage of quadrat squares containing organismPercentage
cover = percentage of ground covered with organism in a quadrat (usually for plants)Pitfall trap = to
collect invertebratesSweep net = to collect invertebrates in long grassesPooter = to collect invertebrates into a
containerTullgren funnel = to collect organisms from soil or leaf litterBaermann funnel = to collect living organisms

from waterconstant changing of abioticconditionsMovement of organismsSampling taken within a smallamount of


timeLimitations of only 1 studyConsideration for safety of organismsDisruption to normal habitatEthics of measuring
wildorganisms
The effect of temperature on thehatching success of brine shrimp
Independent =temperatureDependent =number of hatchedshrimpLight intensitypHsalt contentpresence of chlorine fromtap
wateroxygenconcentrationBrine shrimp eggcysts2 g sea salt for eachtreatmentde-chlorinated waterfor
eachtreatmentbeakersWater baths orincubatorsForcepsBright lightpipetteDecide on a range of temperatures from 5 C to 35
C to be tested.

Place 2 g of sea salt into a 100 cm3 beaker. Add 100 cm3 of de-chlorinated water and stir until the salt completely dissolves.
Label the beaker with sea salt and the temperature at which itwill be incubated. Place a tiny pinch of egg cysts onto a large
sheet of white paper. Wet the piece of graph paper using a fewdrops of salt water. Dab the paper onto the white sheet to pick
up approximately 40 eggs. Use a magnifying glass to count theeggs. Put the paper with the 40 eggs into the beaker (eggsside down). After 3 minutes, use a pair of forceps to gently removethe paper, making sure that all the egg cysts have washed
off into the water. If possible replicate the treatments.

Incubate thebeakers at the appropriate temperatures, controlling exposure to light as far as possible. The next day count the
number of hatched larvae in each of the beakers. To do this, place a bright light next to the beaker. Any larvae will swim
towards the light.Using a fine glass pipette catch the brine shrimps and place them in a small beaker of salt water. Brine
shrimps are verydelicate and care must be taken when handling them. Record the number of larvae that have successfully
hatched at eachtemperature.

Outcome
The majority of the shrimp should hatch at the optimum temperature between 25 and 30C. (optimum at 28C)
. Statstests could be used to show evidence for data.Difference = student t test or mann whitney U
Correlation = spearmans rank
ethics of hatching shrimp underdifferent conditionsuse of animals in experimentseffect of light intensity, may be
adifference in light in each samplefluctuating temperaturesnot accurate salt measurementsmay not have counted exactly
40eggsmay miss seeing some of thebaby shrimpsome eggs may not be viableanymore and wont hatch
DNA gel electrophoresis
selected restrictionenzymesagar gelgel tankelectrical supplymicropipettesDNA sampleLoading dyeUV lightCameraBuffer
solutionDNA restrictionladderMix DNA with desired restriction enzyme and loading dye. Prepare agar and pour into
electrophoresis mould. Once set, fillelectrophoresis tank with buffer solution. Use micropipette to load restriction ladder
into first well then DNA samples cut withrestriction enzyme into the other wells. Connect to electrical supply, turn on
and leave until the dye has moved to the oppositeend of the gel tank. Switch off and disconnect electrical supply. Carefully
remove the gel from the tank and view under UVlight. Take picture if desired.
Outcome
DNA will be separated out through the agar gel, with the heaviest (biggest) DNA strands near the wells and the
lightest (smallest) will be at the opposite end. The DNA restriction ladder can be used as a ruler to measure the size of t
hedifferent fragments.

DNA amplificationusing PCR


ThermocyclerDNA sampleTaq polymeraseNucleotidesprimersDNA sample is placed into tube in thermocycler with
nucleotides, primers and polymerase. Step 1: denaturation = DNA heatedfor 1min at 94C to denature it. This breaks the
H bonds between nucleotides and makes the double stranded DNA, singlestranded.Step 2: annealing = temperature reduced
to 54C.Bonds form between primers and the template strands . This willallow the polymerase enzyme to start to copy the
template.Step 3: extension = carried out at 72C. This is the optimum for taq polymerase enzyme. The bases are placed
in their correctposition, extending the strand from the primer.The amount of DNA doubleseach cycle (steps 1-3) thereforea
considerably amount of copiedDNA can be made for use in DNAfingerprinting etc.35 cycles ( a few hrs) = 34 billioncopies

A2 BIOLOGY CORE PRACTICAL SUMMARY


Name of practicalIndependent &dependentvariablesOther variablesto be
controlledequipment Method and outcome Possible evaluation issues
Observing patternsby Ecological sampling
Random samplingSystemic samplingAbiotic factorse.g. light,temperature, soilwater, humidity,O
2
concentration,pH, aspect, slopeangleGridded QuadratTape measurePoint quadratPitfall trapSweep NetPooterTullgren
funnelBaermann funnelSeveral methods.1
random sampling
= set up grid using tape measure, use random numbers to generate points to place quadrat to collect data.2
systemic sampling
= line transect often used especially to study zonation. A tape measure is laid along several zones to belooked at and
quadrats are used to record data at regular intervals
3 Measuring abundance
Density = presence of organisms per quadratFrequency = percentage of quadrat squares containing organismPercentage
cover = percentage of ground covered with organism in a quadrat (usually for plants)Pitfall trap = to
collect invertebratesSweep net = to collect invertebrates in long grassesPooter = to collect invertebrates into a
containerTullgren funnel = to collect organisms from soil or leaf litterBaermann funnel = to collect living organisms
from waterconstant changing of abioticconditionsMovement of organismsSampling taken within a smallamount of
timeLimitations of only 1 studyConsideration for safety of organismsDisruption to normal habitatEthics of measuring
wildorganisms
The effect of temperature on thehatching success of brine shrimp
Independent =temperatureDependent =number of hatchedshrimpLight intensitypHsalt contentpresence of chlorine fromtap
wateroxygenconcentrationBrine shrimp eggcysts2 g sea salt for eachtreatmentde-chlorinated waterfor
eachtreatmentbeakersWater baths orincubatorsForcepsBright lightpipetteDecide on a range of temperatures from 5 C to 35
C to be tested.

Place 2 g of sea salt into a 100 cm3 beaker. Add 100 cm3 of de-chlorinated water and stir until the salt completely dissolves.
Label the beaker with sea salt and the temperature at which itwill be incubated. Place a tiny pinch of egg cysts onto a large
sheet of white paper. Wet the piece of graph paper using a fewdrops of salt water. Dab the paper onto the white sheet to pick
up approximately 40 eggs. Use a magnifying glass to count theeggs. Put the paper with the 40 eggs into the beaker (eggsside down). After 3 minutes, use a pair of forceps to gently removethe paper, making sure that all the egg cysts have washed
off into the water. If possible replicate the treatments.
Incubate thebeakers at the appropriate temperatures, controlling exposure to light as far as possible. The next day count the
number of hatched larvae in each of the beakers. To do this, place a bright light next to the beaker. Any larvae will swim
towards the light.Using a fine glass pipette catch the brine shrimps and place them in a small beaker of salt water. Brine
shrimps are verydelicate and care must be taken when handling them. Record the number of larvae that have successfully
hatched at eachtemperature.
Outcome
The majority of the shrimp should hatch at the optimum temperature between 25 and 30C. (optimum at 28C)
. Statstests could be used to show evidence for data.Difference = student t test or mann whitney U
Correlation = spearmans rank
ethics of hatching shrimp underdifferent conditionsuse of animals in experimentseffect of light intensity, may be
adifference in light in each samplefluctuating temperaturesnot accurate salt measurementsmay not have counted exactly
40eggsmay miss seeing some of thebaby shrimpsome eggs may not be viableanymore and wont hatch
DNA gel electrophoresis
selected restrictionenzymesagar gelgel tankelectrical supplymicropipettesDNA sampleLoading dyeUV lightCameraBuffer
solutionDNA restrictionladderMix DNA with desired restriction enzyme and loading dye. Prepare agar and pour into
electrophoresis mould. Once set, fillelectrophoresis tank with buffer solution. Use micropipette to load restriction ladder
into first well then DNA samples cut withrestriction enzyme into the other wells. Connect to electrical supply, turn on
and leave until the dye has moved to the oppositeend of the gel tank. Switch off and disconnect electrical supply. Carefully
remove the gel from the tank and view under UVlight. Take picture if desired.
Outcome
DNA will be separated out through the agar gel, with the heaviest (biggest) DNA strands near the wells and the

lightest (smallest) will be at the opposite end. The DNA restriction ladder can be used as a ruler to measure the size of t
hedifferent fragments.
DNA amplificationusing PCR
ThermocyclerDNA sampleTaq polymeraseNucleotidesprimersDNA sample is placed into tube in thermocycler with
nucleotides, primers and polymerase. Step 1: denaturation = DNA heatedfor 1min at 94C to denature it. This breaks the
H bonds between nucleotides and makes the double stranded DNA, singlestranded.Step 2: annealing = temperature reduced
to 54C.Bonds form between primers and the template strands . This willallow the polymerase enzyme to start to copy the
template.Step 3: extension = carried out at 72C. This is the optimum for taq polymerase enzyme. The bases are placed
in their correctposition, extending the strand from the primer.The amount of DNA doubleseach cycle (steps 1-3) thereforea
considerably amount of copiedDNA can be made for use in DNAfingerprinting etc.35 cycles ( a few hrs) = 34 billioncopies
As Biology With Stafford Practical Workbook Marking Schemes (3) (1)
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Marking scheme for AS Biology with Stafford, Unit Three Practical Workbook. Book available at
http://www.amazon.com/gp/aw/s/ref=is_s_?k=AS+A2+Biology+with+stafford
AS Biology with Stafford
.

Unit Three: Practical Workbook


answers
/ Unit three paper BIO7
Page
1

Marking schemes for


Advanced Level Biology
AS Biology with Stafford
Unit Three: Practical Workbook Paper reference: 6BIO7

The book is available at the following link


http://www.amazon.com/gp/aw/s/ref=is_s_?k=AS+A2+Biology+with+stafford
A copy of this marking scheme and many other resources can be downloaded from thefollowing
link http://www.facebook.com/groups/biologywithstafford/

Marking scheme for AS Biology with Stafford, Unit Three Practical Workbook. Book available at
http://www.amazon.com/gp/aw/s/ref=is_s_?k=AS+A2+Biology+with+stafford
AS Biology with Stafford
.

Unit Three: Practical Workbook


answers
/ Unit three paper BIO7

Page
2

Copyright Stafford Valentine ReddenUnauthorized duplication contravenes applicable laws.


Typeset & layouts by
The effect of temperature on thehatching success of brine shrimp
Independent =temperatureDependent =number of hatchedshrimpLight intensitypHsalt contentpresence of chlorine fromtap
wateroxygenconcentrationBrine shrimp eggcysts2 g sea salt for eachtreatmentde-chlorinated waterfor
eachtreatmentbeakersWater baths orincubatorsForcepsBright lightpipetteDecide on a range of temperatures from 5 C to 35
C to be tested.
Place 2 g of sea salt into a 100 cm3 beaker. Add 100 cm3 of de-chlorinated water and stir until the salt completely dissolves.
Label the beaker with sea salt and the temperature at which itwill be incubated. Place a tiny pinch of egg cysts onto a large
sheet of white paper. Wet the piece of graph paper using a fewdrops of salt water. Dab the paper onto the white sheet to pick
up approximately 40 eggs. Use a magnifying glass to count theeggs. Put the paper with the 40 eggs into the beaker (eggsside down). After 3 minutes, use a pair of forceps to gently removethe paper, making sure that all the egg cysts have washed
off into the water. If possible replicate the treatments.
Incubate thebeakers at the appropriate temperatures, controlling exposure to light as far as possible. The next day count the
number of hatched larvae in each of the beakers. To do this, place a bright light next to the beaker. Any larvae will swim
towards the light.Using a fine glass pipette catch the brine shrimps and place them in a small beaker of salt water. Brine
shrimps are verydelicate and care must be taken when handling them. Record the number of larvae that have successfully
hatched at eachtemperature.
Outcome

The majority of the shrimp should hatch at the optimum temperature between 25 and 30C. (optimum at 28C)
. Statstests could be used to show evidence for data.Difference = student t test or mann whitney U
Correlation = spearmans rank
ethics of hatching shrimp underdifferent conditionsuse of animals in experimentseffect of light intensity, may be
adifference in light in each samplefluctuating temperaturesnot accurate salt measurementsmay not have counted exactly
40eggsmay miss seeing some of thebaby shrimpso
The effect of temperature on thehatching success of brine shrimp
Independent =temperatureDependent =number of hatchedshrimpLight intensitypHsalt contentpresence of chlorine fromtap
wateroxygenconcentrationBrine shrimp eggcysts2 g sea salt for eachtreatmentde-chlorinated waterfor
eachtreatmentbeakersWater baths orincubatorsForcepsBright lightpipetteDecide on a range of temperatures from 5 C to 35
C to be tested.
Place 2 g of sea salt into a 100 cm3 beaker. Add 100 cm3 of de-chlorinated water and stir until the salt completely dissolves.
Label the beaker with sea salt and the temperature at which itwill be incubated. Place a tiny pinch of egg cysts onto a large
sheet of white paper. Wet the piece of graph paper using a fewdrops of salt water. Dab the paper onto the white sheet to pick
up approximately 40 eggs. Use a magnifying glass to count theeggs. Put the paper with the 40 eggs into the beaker (eggsside down). After 3 minutes, use a pair of forceps to gently removethe paper, making sure that all the egg cysts have washed
off into the water. If possible replicate the treatments.
Incubate thebeakers at the appropriate temperatures, controlling exposure to light as far as possible. The next day count the
number of hatched larvae in each of the beakers. To do this, place a bright light next to the beaker. Any larvae will swim
towards the light.Using a fine glass pipette catch the brine shrimps and place them in a small beaker of salt water. Brine
shrimps are verydelicate and care must be taken when handling them. Record the number of larvae that have successfully
hatched at eachtemperature.
Outcome
The majority of the shrimp should hatch at the optimum temperature between 25 and 30C. (optimum at 28C)
. Statstests could be used to show evidence for data.Difference = student t test or mann whitney U
Correlation = spearmans rank

ethics of hatching shrimp underdifferent conditionsuse of animals in experimentseffect of light intensity, may be
adifference in light in each samplefluctuating temperaturesnot accurate salt measurementsmay not have counted exactly
40eggsmay miss seeing some of thebaby shrimpsome eggs may not be viableanymore and wont hatc
Another table
DNA gel electrophoresis
selected restrictionenzymesagar gelgel tankelectrical supplymicropipettesDNA sampleLoading dyeUV lightCameraBuffer
solutionDNA restrictionladderMix DNA with desired restriction enzyme and loading dye. Prepare agar and pour into
electrophoresis mould. Once set, fillelectrophoresis tank with buffer solution. Use micropipette to load restriction ladder
into first well then DNA samples cut withrestriction enzyme into the other wells. Connect to electrical supply, turn on
and leave until the dye has moved to the oppositeend of the gel tank. Switch off and disconnect electrical supply. Carefully
remove the gel from the tank and view under UVlight. Take picture if desired.
Outcome
DNA will be separated out through the agar gel, with the heaviest (biggest) DNA strands near the wells and the
lightest (smallest) will be at the opposite end. The DNA restriction ladder can be used as a ruler to measure the size of t
hedifferent fragments.

Another table
DNA amplificationusing PCR

ThermocyclerDNA sampleTaq polymeraseNucleotidesprimersDNA sample is placed into tube in thermocycler with


nucleotides, primers and polymerase. Step 1: denaturation = DNA heatedfor 1min at 94C to denature it. This breaks the
H bonds between nucleotides and makes the double stranded DNA, singlestranded.Step 2: annealing = temperature reduced
to 54C.Bonds form between primers and the template strands . This willallow the polymerase enzyme to start to copy the
template.Step 3: extension = carried out at 72C. This is the optimum for taq polymerase enzyme. The bases are placed
in their correctposition, extending the strand from the primer.The amount of DNA doubleseach cycle (steps 1-3) thereforea
considerably amount of copiedDNA can be made for use in DNAfingerprinting etc.35 cycles ( a few hrs) = 34 billioncopies
Effects of different antibiotics onbacteria
Independen =antibioticDependent =diameter of inhibition zone
Concentrationof antibioticAmount of antibioticDisc sizeBacterialspeciesTemperatureRulerSamples of differentantibiotics
on mastring or filter paperdiscsPetri dishesAgar gelDisinfectantBunsen burnerForcepsMarker penAdhesive
tapeincubatorWash hands. For this practical you will need to work in sterile conditions (aseptic technique) i.e. you will need
to flame theforceps in the Bunsen after every use. Prepare an agar plate seeded with bacteria.Label the Petri dish on the base
at the edgewith your name, the date and the type of bacterium it is inoculated with. Flame the forceps and then use them
to pick up anantibiotic disc or Mast ring. Raise the lid of the Petri dish and place the Mast ring firmly in the centre of the
agar; if individualdiscs are used they will need to be spaced evenly around the dish. Tape the dish securely with two pieces
of adhesive tape (butdo not seal it completely), then keep it upside down at 30Cfor 48 hours. After incubation, look
carefully at the plate but
donot open it

. Where bacteria have grown the plate will look opaque, but where the antibiotics have inhibited growth, clearzones called
inhibition zones will be seen. Measure the diameter of the inhibition zones in millimetres and use this informationto decide
which antibiotic is most effective at inhibiting the growth of the bacterium.
Outcome
: dependent on bacterial species used and antibiotics used. E.g. E.coli is gram negative and not often susceptible topenicillin
which is effective mainly with gram positive species. The larger the inhibition zone, the more effective the antibioticagainst
that species. Ensuring that the discs areplaced evenly on the Petri dishHaving good aseptic techniqueto prevent
plate contaminationAge of antibiotic, if the antibioticused is out of date it is likely tobe less effectiveRepeatsAccuracy of
incubationtemperature and time
Another table
Measuring the rateof oxygen uptake
No of organismsTemperatureTimeAmount of soda limeRespirometerSoda limeColoured liquid5g Organisms
e.g.maggots,germinating peas,woodliceCotton woolStop clockMarker penPlace 5g of organism (maggots) into the tube and
replace the bung. Introduce a drop of dye into the glass tube. Opentheconnection (three-way tap) to the syringe and move
the fluid to a convenient place on the pipette (i.e. towards the end of thescale that is furthest from the test tube). Mark the
starting position of the fluid on the pipette tube with a permanent OHT pen.Isolate the respirometer by closing the
connection to the syringe and the atmosphere and immediately start the stop clock.Mark the position of the fluid on the
pipette at 1 minute intervals for 5 minutes. 6. At the end of 5 minutes open theconnection to the outside air. Measure the
distance travelled by the liquid during each minute (the distance from one mark tothe next on your pipette).If your tube does
not have volumes marked onto it you will need to convert the distance moved into

volume of oxygen used. (Remember the volume used = r2 distance moved, where r = the radius of the hole in the
pipette.)
Record your results in a suitable table. Calculate the mean rate of oxygen uptake during the 5 minutes.
Outcome
: Oxygen molecules are absorbed by the organism and used in respiration. The same number of carbon dioxidemolecules are
released but these are absorbed by the soda lime. This reduces the pressure inside the test tube (fewer molecules =lower
pressure). Atmospheric pressure pushes the liquid along the tube, until the pressure in and outside the tube is equal.Oxygen
is the final electron acceptor, and it eventually combines with hydrogen to make water. The carbon dioxide comes fromthe
carbon dioxide released in the link reaction and the Krebs cycle as the carbohydrate is broken down.Simple respirometer

disadv
. =does not allow you to reset; itneeds a control tube usedalongside it; no scale someasurements likely to be lessaccurate.
Adv
= very simple toset up; minimal number of connections makes a good sealeasier to obtain. U-tube respirometer

disadv

. =tendency for the connections toleak in elderly school/collegemodels (making the equipmentuseless); expense.
Adv
. = doesnot need to have an additionalcontrol as the second tubebalances out the effects of changes in temperature
oratmospheric pressure; the syringeallows you to move the liquid inthe U to reset the apparatus.
Another table
Effects of exerciseon tidal volume and breathing rate
SpirometerKymographDisinifectantEye protectionSoda limeThe general principle behind a spirometer is simple. It
is effectively a tank of water with an air-filled chamber suspended in thewater. It is set up so that adding air to the chamber
makes the lid of the chamber rise in the water, and removing air makes itfall. Movements of the chamber are recorded
using a kymograph (pen writing on a rotating drum). Tubes run from thechamber to a mouthpiece and back again. Breathing
in and out through the tubes makes the lid of the chamber fall and rise.The volume of air the person inhales and exhales can
be calculated from the distance the lid moves. The apparatus can becalibrated so that the movement of the lid corresponds to
a given volume. A canister containing soda lime is inserted betweenthe mouthpiece and the floating chamber. This absorbs
the CO2 that the subject exhales. In which direction will the pen movewhen the subject inhales. After calibration, the
spirometer is filled with oxygen. A disinfected mouthpiece is attached to thetube, with the tap positioned so that the
mouthpiece is connected to the outside air. The subject to be tested puts a nose clipon, places the mouthpiece in their mouth
and breathes the outside air until they are comfortable with breathing through thetube. Switch on the recording apparatus
and at the end of an exhaled breath turn the tap so that the mouthpiece is connectedto the spirometer chamber. The trace will
move down as the person breathes in. After breathing normally the subject shouldtake as deep a breath as possible and then
exhale as much air as possible before returning to normal breathing. See traceexample below.
Outcome

: The tidal volume is the volume of air breathed in and out in one breath at rest. The tidal volume for most adults isonly
about 0.5 dm3. Vital capacity is the maximum volume of air that can be breathed in or out of the lungs in one forcedbreath.
Breathing rate is the number of breaths taken per minute. Minute ventilation is the volume of air breathed into (andout of)
the lungs in one minute. Minute ventilation = tidal volume rate of breathing (measured in number of breaths perminute).
Some air (about 1 dm3) always remains in the lungs as residual air and cannot be breathed out. Residual air preventsthe
walls of the bronchioles and alveoli from sticking together. Any air breathed in mixes with this residual air.

Another table
Another table
nvestigatinghabituation to astimulusIndependent variable = number of pokesDependent variable= retraction time
Replicationusing snails of approx samesize and ageEqual handlinghistoryDrying out1 giant African landsnail1 dampened
cottonwool budClean firm surfaceStop watchCollect one giant African land snail, and place it on a clean, firm surface.
Allow the snail to get used to its new surroundings fora few minutes until it has fully emerged from its shell. Dampen
a cotton wool bud with water. Firmly touch the snail betweenthe eye stalks with the dampened cotton wool bud and
immediately start the stopwatch. Measure the length of time betweenthe touch and the snail being fully emerged from its
shell once again, with its eye stalks fully extended. Repeat the procedure instep 3 for a total of 10 touches, timing how long
the snail takes to re-emerge each time. Record your results in a suitable table.Present your results in an appropriate graph.
Outcome
: spearmans rank stats test to look for correlation in data.
There is a negative correlation


as the number of stimuliincrease the time taken for the snail to re-emerge decreases. Students should make a reference to the
data. With repeatedstimulation, Ca2+ channels in the presynaptic membrane become less responsive. Less Ca2+ crosses the
membrane into thepresynaptic (sensory) neurone. As a result less neurotransmitter is released into the synaptic cleft. This
means that an actionpotential across the postsynaptic membrane is less likely. Fewer action potentials are produced in
the postsynaptic motorneurone so less of a response is observed.Snails already handled beforethe experiment may not react
inthe same wayDetermining when a snail hasfully emergedLack of moisture may encouragesnail to stay more in its
shellMeasuring eye stalk lengthinstead

: Mohamed Sobir
Cover designed by
: Mohamed Sobir
Printed in Maldives by

: Copier Repair
Published by
: Author publisher All rights reserved. No part of this publication may beReproduced, stored in a database or retrieval
system, or transmitted in anyform or by any means, electronic,mechanical, photocopying, recording,or otherwise, without
the prior written permission of the author
ISBN:
978-81-910705-2-1
Note: A few graphs have been drawn to give students an idea on how to find the scaleand plot range or variability.
Other graphs have not been drawn as students need topractice the graph drawing. Do post your graphs on my FB
group and I can give afeedback on the graphs.Any other queries on any aspect of the practical paper can be clarified
on my FB grouphttp://www.facebook.com/groups/biologywithstafford/I have also included additional notes on
referencing, citation or bibliography at the endof the document. Evaluation of references is also dealt with in detail.
Cheers and all the best.
Stafford Valentine Redden
(M.A; M.Sc.; M.Ed.; (Ph.D))Head of Department (Biology)Villa International School,
Male, Republic of Maldives
Email: staffordv@yahoo.comMobile: +960 7765507

Observing Mitosis
1.

Heat 2cm3 of 1 mole HCl acidin a 60oC waterbath.

2.

Cut off 1-2cm garlic root tips.

3.

Put the garlic root tips in a watch glass containing 2cm3 of acetic alcohol for 12 minutes.

4.

Remove the tips from the acetic acid and place them into a different watch glass containing5cm3 of ice cold distilled
water.

5.

After 5 minutes remove themand leave them to dry.

6.

Then place the tips into thewarmed HCl acid for 5 minutes.

7.

Repeat 1-6 to make the tips even more fragile.

8.

Transfer a tip to a microscope slide and then cut 5mm off the end of the tip.

9.

Macerate using a mounted needle.

10. Stain using 1 drop of toludine blue and leave root tips for 2 minutes.
11. Add coverslip and blot with filter paper.
12. View under a microscope and identify the stages of mitosis.

(Overview of treatment to 1-2cm tips: 2cm3 acetic alcohol -> 5cm3 ice cold water -> dry -> 2cm3 60oC HCl acid > repeat -> Cut 4-5mm tip off root tip ->macerate using mounted needle -> stain with toludine blue -> blot with filter
paper -> view)

Source: LostWacky's
Mint or Garlic in Toothpaste
1. Seed agar plates using aseptic techniques.
2. Crush 3g of mint leaves using a pestle and mortar in 10cm3 of methylated spirit.
3. Pipette 0.1cm3 of the mint leaf solution onto a paper disc.
4. Repeat 1-3 using garlic leaves instead of mint leaves.
5. Allow the discs to dry for 10 minutes.

6. Using sterilised forceps place the discs onto a petri dishes.


7. Use a blank disc as a control in the petri dish.
8. Tape the lid onto the petri dish, leaving a sufficient air supply so that dangerous anaerobes do not grow.
9. Incubate the petri dish for 24 hours at 25oC.
10. Measure the diameter of the rings of affect around the different discs and record the data in a table.
11. Repeat 1-10 at least 3 times and calculate mean averages.
12. Create a graph in order to better see the differences between the antibacterial effects of mint and garlic.
13. Larger diameters of the rings of affect suggest more antibacterial strength.
For a more in-depth (not tailored to SNAB) version of this experiment, see: Garlic and Mint Experiment

McCartney Bottle

Source: Aliimg.com
Totipotency and Tissue Culture
1.

Acquire seeds that are starting to unfold their cotyledons.

2.

Cut the tops of the seeds just below the shoot apex usingsharp scissors.

3.

Place the stem of the newexplants into agar gel inMcCartney bottles.

4.

Cover the McCartney bottles with cling film and place on a sunny windowsill.

Factors that may affect the results:

Agar gel may become contaminated with pathogens (which kill the cotyledons) because of the nutritious and moist
environment it provides.
The wrong part of the plant may be cut off and placed into the agar gel.

Plant cross section (should remember from previous unit)


Source: BBC

The Strength of Plant Fibres


1.

Soak plant material in water for at least one week to soften the fibres for easier extraction.

2.

Ret the plant material for its fibres.

3.

Clamp a fibre between twoclamp stands and add mass to the centre of the fibre until it snaps (record the weight at
which it does so).

4.

Repeat at least 3 times and calculate the average.

5.

Repeat using other plant fibres

6.

Plot data in a bar graph to compare fibre strengths.

7.

Ensure that the lengths of the fibres are equal each time.

8.

Ensure that the plants are the same age at the time of extraction as older fibres will be more brittle than younger
counterparts.

A tomato plant deprived of magnesium.


Source: Plantphys.net
Investigating Plant Mineral Deficiencies
1.
2.

Prepare bottles of varying mineral content starting from a control of distilled water(lacking all nutrients) and ending
with a solution containing all nutrients (with no potassium, no phosphorus, no magnesium etc. in other bottles).
Cover bottle openings with foil and then perforate the centre of the foil lids.

3.

Place a plant of the same species in each bottle by putting its roots through the opening that was perforated so that the
roots may absorb the solution.

4.

Place on a sunny windowsill.

5.

Record observations.

Note: see photo below for major roles of each nutrient.

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Major Functions of Mineral Deficiencies

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