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SAPS Biotechnology Scotland Project
Laboratory Sessions
Biotechnology Summer School 2002
Aim of practical sessions
To provide hands-on experience of a range of experimental work which:
enlivens teaching of molecular biology, biotechnology, genetics & plant science
is appropriate for 5-14 through to AH
supports curriculum
aids pupil understanding
Practical Sessions
Monday
Fruit Full of DNA
Protein Power
The Wonderful Wizardry of Finding a Gene
Virus Detectives
Friday
Fruit Full of DNA - Curriculum
Simple extraction of DNA from kiwi & strawberry
5 – 14 Environmental Studies: Science
Outline the principles of modern biotechnology and explain its significance now and for the future
Explain the role of chromosomes and genes in inheritance
Simple extraction of DNA from kiwi & strawberry
Standard Grade Biology
Inheritance: Sub-Topic B, What is Inheritance?
Unit 1 Living Cells: (b) Factors affecting variety in a species: Genetics
Intermediate 2 Biology
Fruit Full of DNA - Curriculum
Fruit Full of DNA - method
Keep materials cool where indicated
Squash fruit in a self-seal plastic bag to mechanically break up cells
Add washing up liquid, salt and heat to 60C for 15 mins to disrupt cellular membranes
Filter to remove cell debris
Precipitate DNA by adding cold methanol
Practical session (a)
Prepare gel for Protein Electrophoresis: Page 13, ‘Preparing the Gel’; Instructions Nos 1, 4 - 6
Squash fruit in a self-seal plastic bag to mechanically break up cells
add washing up liquid, salt and heat to 60C for 15 mins to disrupt cellular membranes
Protein Power
DNA Extraction
Protein Power
Extraction & electrophoresis of protein
from a variety of sources
Protein Power
Gel Electrophoresis
negative charge conferred on molecules
gel is porous
molecules travel through gel according to size when electrical current applied
smaller travel faster
Protein Power
Protein within subject specifications
Nutrition – demonstrate different foods contain proteins
Structure and function – demonstrate proteins of different sizes
Standard Grade Biology
Animal Survival: Sub-Topic a, The need for food
Unit 3: (b) Animal Physiology: Mammalian nutrition
Intermediate 2 Biology
Protein Power - Curriculum
Higher Biology
Unit 1, Cell Biology: (d) Synthesis and release of proteins
Unit 1, Cell Function and Inheritance: (b) Protein synthesis
Higher Human Biology
Protein Power - Curriculum
Higher Biotechnology
Unit 1, Microbiology: (b) Microbial metabolism
Unit 1, Cell and Molecular Biology: (b) Structure and function of cell components
Investigation
Advanced Higher Biology
Protein Power - Curriculum
Protein Power - Method
Extract the protein Cut sample – !SIZE MATTERS! – small is beautiful
Mash sample
Add blue marker dye (Laemmli buffer) – it contains
Blue dyeGlycerolDetergent Mix by flicking tube
Denature proteins by placing in boiling water bath
Cool – leave on ice
Protein Power - Method
Load the gel Gel – pure agarose (3%)
Cover with TB Buffer (conducts electricity)
Gently remove comb from gel
Load wells – clean tip for each sample
Work close to electrophoresis tank
Protein Power - Method
Place in large electrophoresis tank
Run the gel
Fit one electrode at each end of the tank
Practical session (b)
DNA Extraction
Filter DNA mixture
Precipitate DNA by adding meths
Prepare protein samples (2 per pair)
Load gels (6 samples per pair)
Run gels (large tank)
Protein Power
Wizard Genes
Make and pour agar gel
Electrophoresis of mixtures of food dyes
Wizard Genes
Importance of gel electrophoresis:
Gene identification, manipulation & sequencing
Protein isolation and identification
Aim of practical:
To introduce idea of separating mixtures and using bands to assist in identification
Electrophoresis of mixtures of food dyes
Wizard Genes - curriculum
5 – 14 Environmental Studies: Science
Outline the principles of modern biotechnology and explain its significance now and for the future
Explain the role of chromosomes and genes in inheritance
Electrophoresis of mixtures of food dyes
Wizard Genes - curriculum
Standard Grade Biology
Inheritance: Sub-Topic B, What is Inheritance?
Unit 1 Living Cells: (b) Factors affecting variety in a species: Genetics
Intermediate 2 Biology
Cross-curricular activities
Wizard genes - method
Prepare agar gel & allow to set
Load wells with ‘chromosome extract’
Insert electrodes and attach crocodile clips
Wire electrophoresis tank to batteries
Run for 30 minutes
Pour off water and examine bands on gel
Identify powers possessed by each wizard
Practical session (c)
Wizard Genes
Practise using micro-pipettors
Load wells and run gels
Stop electrophoresis, wash gels & soak for an hour. Remember to rinse after half an hour
Protein Power
Discard washing water
Stain with Coomassie Blue Stain
Protein Power
Protein Power - Method
Staining the proteins
Discard electrodes and pour away buffer
Wash with deionised water (soak for 1 hour)
Shake stain bottle well and pour 12cm3 over gel
Leave for 24 hours and wash
Results best seen 24 – 48 hours after staining
See you tonight
for visit to
Mary King’s Close
Biotechnology Summer School 2002