Unit 3 Flashcards
How might you carry out the Benedicts test?
Create solution. Add equal amount of reagent to solution. Shake + heat in water bath. Partially quantitative.
What is the test for reducing sugars?
Benedicts test
What is the test for non-reducing sugars?
Modified Benedicts test
How might you carry out the modified Benedicts test?
Hydrolyse by boiling in dilute HCL. Once cooled, neutralise with sodium hydrogen carbonate. Check with pH paper and repeat Benedicts test.
What are the results of the Benedicts test?
Blue - green - yellow - orange - brick red precipitate.
What is the test for proteins?
Biuret test. Detects presence of peptide links.
How might you carry out the test for proteins?
Create solution. Add potassium hydroxide to substance, then add copper sulphate + shake.
What are the results of the test for proteins?
Blue - lilac
What is the test for lipids?
Emulsion test
How might you carry out the test for lipids?
Add sample to ethanol + shake, allow to settle, empty liquid into test tube w/water.
What are the results of the test for lipids?
Clear/colourless - milky white emulsion.
What is the test for glucose?
Clinistix
How might you carry out the test for glucose?
Dip stick into substance and wait for a few minutes.
What are the results of the test for glucose?
It will change colour and the level can be found by checking the label on the bottle.
What is paper chromatography used for?
Identifying different amino acids. More soluble substance travels further.
How might you carry out paper chromatography?
Cut paper to size, add solvent to tank + saturate atmosphere (Lid on), draw base line, add spot of solution + mark with x, repeat x5 for conc spot, place in tank with line above solvent, Draw line on solvent front before reaches top of paper (solvent travels up with dissolved AA’s). Allow to dry + spray with ninhydrin in fume cupboard. Heat gently. Circle spots + calculate Rf.
How do you find the Rf value of an amino acid you tested using paper chromatography?
Distance moved by spot/distance moved by solvent..
What is the test for starch?
Iodine
How might you carry out the test for starch?
Add a few drops of iodine to the food sample.
What are the results of the test for starch?
Yellow/brown to blue/black.
Will hydrophilic amino acids prefer to stay in the stationary phase or the mobile phase?
What about hydrophobic amino acids?
What would their Rf values be?
Stationary phase, lower.
Mobile phase, higher.
How might you determine the effects of pH on enzyme activity? Which pH works best?
Jelly cubes in different pH buffers with a protease in a water bath of optimum temp (35c). Test percentage transmission with a colorimeter. The pH with the lowest percentage transmission as the dye is released and less light can pass through. Control temp, enzyme vol + conc + substrate vol + conc.
Where is trypsin found? What is its optimum pH?
Small intestine. pH 7
How might you determine the effects of temperature on enzyme activity? Which temperature works best?
Mix starch + amylase at required temp + place at 5 diff temps in thermostatically controlled water bath with iodine. Use a colorimeter to measure their percentage transmission. The temperature with the highest percentage transmission is the optimum temperature as the starch is not present according to the iodine. pH, enzyme volume + conc + substrate vol + conc must be controlled.
How does a colorimeter work?
Measures change in intensity of light as it passes through a solution. They can record the amount of light absorbed by solution (absorbance) or amount that passes through (transmission). The light that isn’t absorbed passes onto a photo-sensitive light + electrical current in converted into digital signal. Must be calibrated by blank solution (100% transmission)
What is a calibration curve?
A graph of known quantities. Allows colorimeter readings to be expressed in substrate concs. Used for calcing spec quantities. Start with known conc of starch, make range of concs, measure % transmission for each value + plot graph with transmission of y axis + conc on x axis.
How might you immobilise lactase to create lactose free milk?
Mix alginate + lactase, add food colouring, place small beads in calcium chloride with a syringe, then allow to harden and pass milk through beads. Test with clinistix to ensure glucose is present so lactose has been broken down.
How might we investigate the distribution of catalase in soaked pea and temperature on this reaction?
Test for catalase by crushing a soaked pea and adding hydrogen peroxide. Test seed coats and cotyledons separately for catalase. Boil each pea and place at different temperatures.
What is alcoholic fermentation? Equation?
In anaerobic conditions yeast cells break down sugar into ethanol and carbon dioxide.
Glucose -> Ethanol + Carbon Dioxide + Energy
How might we test the specificity of enzymes?
Add yeast to different solutions of sugars. Test how the measurement of liquids changes.
How might you prepare a slide to look at in a microscope?
Place a small amount of the substance onto a slide, add a drop of water/iodine onto it and add a cover slip.
How do you use a stage micrometer?
You must first calibrate the stage scale with the eyepiece scale so that the size of the divisions in the eyepiece are known at high and low power.
How do you calibrate an eyepiece graticule?
Align the top of both scales. Count the number of divisions on the eyepiece graticule that add up to a fixed number of divisions on the stage micrometer. You know it’s the eyepiece as when you move focus knob, it doesn’t change.
Why might a student blot dry potato cylinders before weighing after having them placed in a sucrose solution?
To ensure that the change in weight was due to changes in potato mass/osmosis, not due to outside liquid.
On a graph of percentage gain in mass vs percentage sucrose solution in an experiment on potato cylinders, how might you find what the concentration of cell sap was?
The point of equilibrium where osmosis is not occurring and there is no change in mass. This is the natural state of the potato so is the sap conc.
How do you know what the average solute potential of an onion cell is when on a graph of solute potential v plasmolysed cell percentage?
The point where 50% of cells are plasmolysed shows the average solute potential of onion cells which is equal to sucrose solute potential. Solute potential of sucrose at that conc can be calculated from conversion table.
How might you perform a root tip squash?
Grow beans in tray for 7-8 days, place section of lateral white root which is actively dividing in boiling tube with acetic orcein stain + heat @ 60 for 30 mins to soften + stain tissue. Cut off very end of root tip which is dividing. Then prepare a microscope slide and tap on cover slip or roll over with fist to squash root to get a single layer of cells. View and find out how long each stage of mitosis occurs for.
What is the formula to find out how long each stage of mitosis took in a root tip squash?
Percentage of cells at stage x 1440 minutes = number of minutes per that stage.
What is a block diagram?
Diagram which is used to show tissue layers, not individual cells.
How do you draw a block diagram?
Accurate rep, proportionality, smooth lines, pencil, ruled line for label,
What does annotate mean?
Label cell and give a short description of its role.
What does hydrogencarbonate indicator test for?
CO2
What are the colours of hydrogencarbonate indicator:
Less CO2
Normal CO2
High CO2
Purple, orange-red, yellow.
What is a simple respirometer used for?
Measure RQ of germinating seeds or small animals (mealworms + woodlice) Measures changes in gas pressure inside apparatus.
Describe how a respirometer experiment can be carried out.
Potassium hydroxide solution in tube to absorb CO2, equal volume of water in other tube to ensure temp/pressure changes act equally on both sides of manometer + cancel out. Wire basket into KOH tube, both tubes in water bath at 20c with taps closed for 15 mins. As respire, take up O2 + give out CO2 which is absorbed causing pressure to fall so fluid of manometer rises. Rise is recorded + O2 volume used is calculated by divide mean rise by time taken. Syringe used to reset fluid. KOH then removed + replaced with water + experiment repeated giving measure of CO2 produced. If CO2 produced = O2 used, no change in manometer levels. If more CO2 produced than O2 used, liquid in KOH tube falls.
What is a respirometer?
Device used to measure rate of respiration of living organism by measuring rate of exchange of O2/CO2. When respiring, it uses O2 + produces CO2 + total gas volume stays same. KOH absorbs CO2 + so volume gas decreases, decreasing pressure. Rate pressure falls = respiration rate.
Why should temp be controlled in a respirometer experiment?
Affects resp rate + change gas volume due to expansion or contraction.
How would you make a 0.1% solution from a 1% starch solution?
1cm3 of starch solution and 9cm3 of distilled water.
State 2 precautions for accuracy when making serial dilutions.
Use a clean pipette + ensure the suspension is thoroughly mixed.
Describe how serial dilutions can be used to create a calibration curve with a colorimeter.
Use colorimeter to find percentage transmission of each dilution + record. Create a graph of % transmission against starch conc. Each solution less concentrated than one before by set dilution factor. (e.g. DF 10, each 10x less conc)
Why might systematic sampling be used on a rocky shore to sample seaweed?
There is gradation/zonation (of conditions) up the shore/
an environmental gradient exists/area to be sampled is not homogenous.
Why might students avoid placing transect tape over rock pools when sampling seaweed?
It isn’t representative of the shore.