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: wenlivens teaching of molecular biology, biotechnology, genetics & plant science wis appropriate for 5-14 through to AH wsupports curriculum waids pupil understanding

Practical Sessions Monday wFruit Full of DNA wProtein Power wThe Wonderful Wizardry of Finding a Gene wVirus Detectives Friday

Fruit Full of DNA - Curriculum Simple extraction of DNA from kiwi & strawberry 5 – 14 Environmental Studies: Science wOutline the principles of modern biotechnology and explain its significance now and for the future wExplain the role of chromosomes and genes in inheritance

Simple extraction of DNA from kiwi & strawberry Standard Grade Biology wInheritance: Sub-Topic B, What is Inheritance? wUnit 1 Living Cells: (b) Factors affecting variety in a species: Genetics Intermediate 2 Biology Fruit Full of DNA - Curriculum

Fruit Full of DNA - method wKeep materials cool where indicated wSquash fruit in a self-seal plastic bag to mechanically break up cells wAdd washing up liquid, salt and heat to 60C  for 15 mins to disrupt cellular membranes wFilter to remove cell debris wPrecipitate DNA by adding cold methanol

Practical session (a) wPrepare gel for Protein Electrophoresis: Page 13, ‘Preparing the Gel’; Instructions Nos 1, wSquash fruit in a self-seal plastic bag to mechanically break up cells wadd 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 wnegative charge conferred on molecules wgel is porous w molecules travel through gel according to size when electrical current applied wsmaller travel faster

Protein Power Protein within subject specifications wNutrition – demonstrate different foods contain proteins wStructure and function – demonstrate proteins of different sizes

Standard Grade Biology wAnimal Survival: Sub-Topic a, The need for food wUnit 3: (b) Animal Physiology: Mammalian nutrition Intermediate 2 Biology Protein Power - Curriculum

Higher Biology wUnit 1, Cell Biology: (d) Synthesis and release of proteins wUnit 1, Cell Function and Inheritance: (b) Protein synthesis Higher Human Biology Protein Power - Curriculum

Higher Biotechnology wUnit 1, Microbiology: (b) Microbial metabolism wUnit 1, Cell and Molecular Biology: (b) Structure and function of cell components wInvestigation Advanced Higher Biology Protein Power - Curriculum

Protein Power - Method Extract the protein wCut sample – !SIZE MATTERS! – small is beautiful wMash sample wAdd blue marker dye (Laemmli buffer) – it contains eBlue dyef eGlycerolf eDetergentf wMix by flicking tube wDenature proteins by placing in boiling water bath wCool – leave on ice

Protein Power - Method Load the gel wGel – pure agarose (3%) wCover with TB Buffer (conducts electricity) wGently remove comb from gel wLoad wells – clean tip for each sample wWork close to electrophoresis tank

Protein Power - Method wPlace in large electrophoresis tank Run the gel wFit one electrode at each end of the tank

Practical session (b) DNA Extraction wFilter DNA mixture wPrecipitate DNA by adding meths wPrepare protein samples (2 per pair) wLoad gels (6 samples per pair) wRun gels (large tank) Protein Power Wizard Genes wMake and pour agar gel

Electrophoresis of mixtures of food dyes Wizard Genes Importance of gel electrophoresis: wGene identification, manipulation & sequencing wProtein isolation and identification Aim of practical: wTo 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 wOutline the principles of modern biotechnology and explain its significance now and for the future wExplain the role of chromosomes and genes in inheritance

Electrophoresis of mixtures of food dyes Wizard Genes - curriculum Standard Grade Biology wInheritance: Sub-Topic B, What is Inheritance? wUnit 1 Living Cells: (b) Factors affecting variety in a species: Genetics Intermediate 2 Biology Cross-curricular activities

Wizard genes - method wPrepare agar gel & allow to set wLoad wells with ‘chromosome extract’ wInsert electrodes and attach crocodile clips wWire electrophoresis tank to batteries wRun for 30 minutes wPour off water and examine bands on gel wIdentify powers possessed by each wizard

Practical session (c) Wizard Genes wPractise using micro-pipettors wLoad wells and run gels wStop electrophoresis, wash gels & soak for an hour. Remember to rinse after half an hour Protein Power wDiscard washing water wStain with Coomassie Blue Stain Protein Power

Protein Power - Method Staining the proteins wDiscard electrodes and pour away buffer wWash with deionised water (soak for 1 hour) wShake stain bottle well and pour 12cm 3 over gel wLeave for 24 hours and wash wResults best seen 24 – 48 hours after staining

See you tonight for visit to Mary King’s Close Biotechnology Summer School 2002