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