ACTIVITAT ENZIMÀTICA DE LA CATALASA EN ELS DIFERENTS TEIXITS ANIMALS I VEGETALS.

1. INTRODUCCIÓ

Valorem de forma quantitativa l'activitat enzimàtica de la Catalasa del fetge del pollastre. Dividirem la pràctica en dues parts. En la primera observarem la diferència d'activitat de la catalasa en diferents teixits animals i vegetals. Per aquesta part utilitzarem la patata, el tomàquet i el fetge de pollastre. En la segona part veurem l'influència de determinants factors en l'activitat enzimàtica.

La catalasa és un enzim present en els peroxisomers de les cèl·lules animals i vegetals que s'encarrega d'eliminar l'aigua oxigenada que es forma en algunes reaccions del metabolisme. La reacció química és la següent:


                                          H2O2----------------------> 2H2O + O2

2. MATERIAL

- Fetge i cor
- Patata, tomàquet i pastanaga
- Tub d'assaig
- Termòmetre
- Pinces
- Bisturí
- Aigua oxigenada al 3%

3. PROTOCOL

1.- Experiment 1.

- Tallem 2 trossos de patata i el tomàquet que pesin més o menys el mateix (mateixa mida) (1cm3 aprox.)

- També tallarem un tros de fetge de la mateixa mida i pes.

- Ho posarem en tres tubs:

1er tub: patata + 5mL aigua destil·lada
2on tub: tomàquet + 5mL aigua destil·lada
3er tub: pastanaga + 5mL aigua destil·lada
4art tub: fetge + 5mL aigua destil·lada
5è tub: cor + 5 mL aigua destil·lada

- Posarem 2mL d'aigua oxigenada i marcarem l'alçada que assoleixen les bombolles en cada tub. Mesurarem aquesta alçada en mm.

4. PREGUNTES

- Variable dependent i independent?
Dependent: L'alçada de les bombolles
Independent: Diferents teixits
- Problema que vol investigar?
Quin teixits presenta més activitat de la catalasa?
- Explicació del resultasts:
Els texts animals presenten més activitat que els vegetals.

5. RESULTATS

Teixit	Alçada bombolles (mm)
Patata	45mm
Tomàquet	45mm
Pastanaga	47mm
Fetge	98mm
Cor	60mm

Experiment 2.

1er tub: Tros de teixit animal a temperatura ambient

2on tub: Tros de teixit animal amb 10 mL d'HCL al 10%

3er tub: Tros de teixit animal congelat

4at tub: Tros de teixit animal bullit

5è tub: Tros de teixit submergit en una dissolució saturada de NaCl

- Afegirem 2mL d'aigua oxigenada i anotarem l'alçada de les bombolles

1. RESULTATS

Tubs	Alçada de bombolles (mm)
Tub 1	60mm
Tub 2	41mm
Tub 3	58mm
Tub 4	44mm
Tub 5	42mm

L21: MITOSI

1. OBJECTIVES

- Find the phases of cell divisions onion.

2. MATERIAL

- Onion
- Orceine A and B
- Dropper
- Whatch glass
- Forceps
- Beaker
- Bunser burner
- Lighter
- Microscope

3. PROCEDUCE

1. A week ago we left an onion in a beaker with some water, (only the tip of the onion touched it) so its roots will grow so we can see the process of mitosis.
2. To start our experiment we took the onion and cut the tip of a root and put it in the watch glass.
3. Then with the dropper we took the orceine A and put some drops on the root and we took the watch glass with the wooden forceps and put it on the bunsen burner so the orceine and the root would heat. Some acid fumes began to evaporate. We had to be careful to not to burn the root so the watch glass could not be too hot, we should be able to touch it with our hand!!
4. After that we took the root with the forceps and put it on a slide and added a couple of drops of orceine B, we waited a couple of minutes.
5. Then with the scalpel we cut 3mm leaving the tip, and always knowing where the tip is. 
6. Finally we added a coverslip and used the squash method so we could observe the cells on the microscope.

L20: PHOTOSYNTHESIS

1. OBJECTIVES

- Relate the light intensy with the photosynthesis process.
- Measure the rate of photosynthesis.
- Identify the products of the process and the variables that can affect it.

2. MATERIAL

- Algae (Elodea)
- 600 mL beaker
- Test tube
- Tap water
- Funnel
- Light souce
- Metric ruler

3. PROCEDUCE

1. First we assigned the different distances to do the experiment and compare the results to each group.
2. We took the 600 ml beaker and placed 7 g of an algae under a clear funnel inside the beaker. The funnel was raised off the bottom on pieces of blue-tack to allow unhampered diffusion of CO2 to Elodea. 
3. We didn't have sodium bicarbonate so we filled the beaker with tap water, the algae and the funnel should be completely under the water.
4. Then we filled a test tube with tap water and placed the thumb over the end of the test tube. We turned the test tube upside down taking care that no air enters and no water comes out and we put this test tube over the end of the funnel (the skinny part)
5. We marked the level of the water on the surface of the test tube with a marker pen.
6. Each group placed the preapartion close to a light source, each group placed the preparation in a different distance 5, 10, 20 and 25 cm, and one with no light source.
7. We also measured the temperature.
8. Finally we left this preparation for and hour and a half. After this time we measured the difference of gas accumulation on the top of the test tube. 

4. QUESTIONS 

1. Identify the dependent and independent variable of this experiment?

The dependent variable is the gas accumulation and the independent is the distance of the light.

2. Using the data from your results prepare a graph and describe what happened to amount of gas in the test tube.

 Andrea's blog.

3. How much gas produced in the test tube after 1 hour? And 1 hour and half?

4. Write the photosynthesis equation. Explain each part of equation. Which substances are produced by photosynthesis? Wich gas is produced that we need in order to live? 

6 CO2 + 6 H2O--------> C6H12O6+ 6 O2

O2------------>CO2

L19: CELLS ORGANELLES

 

Tomato Chromoplasts: 4 x 10= 40X

  

Potato amyloplasts, stained with lugol: 10 x 40 = 400X

Potato amyloplasts, stained with lugol: 10 x 100 = 1000X

Chromoplasts red cabbage: 15 x 10 = 150X

Chromoplasts red cabbage; 15 x 40= 600X

Stoma of a red cabbage; 15 x 40= 600X

Carrot Chromoplasts : 100X

Chloroplasts of Vallisneria

L18: LIFE IN A DROP OF WATER

In this picture we can see flagel, that moves throught water.
The flagel moves very quickly. Thanks to its movement, causes current of water.

We can see the video: http://www.youtube.com/feature=player_embedded#t=30


In this pic

L17: GRAM STAINING

1. OBJECTIVES

- Differenciate yogurt bacteria.
- Relate the staining procedure with the structure of the cells.

2. MATERIAL:

- 1 Slide
- 1 Cover slip
- Tongs
- Needle
- Gram stain: crystal violet, iodine and safranin.
- Descolorize reagent: ethanol 96%
- Microscope
- Yogurt

3. PROCEDURE

- PROKARIOTIC CELL OBSERVATIONS: GRAM STAINING

1. Prepare a heat-fixed sample of the bacteria to be stained.
2. Cover the smear with crystal violet for an exposure of 1 min.
3. Rinse with distilled water.
4. Apply Iodine solution for 1 min.
5. Rinse the sample with distilled water.
6. Decolorize using ethanol. Drop by drop until the purple stops flowing. Wash immediately with distilled water.
7. Cover the sample with the safrain stain for an exposure time of 45 seconds.
8. Rinse the sample with distilled water.
9. Gently dry the slide with paper.

Gram-negative: stain pink of reddish color.
Gram-positive: stain purple color.

4. RESULTS AND OBSERVATIONS

L16: EPIDERMIS CELLS

1. OBJECTIVES

- Identify the shape of epidermis cells.
- Identify and explore the parts of stoma.
- Measure dimensions of the entire cell and stoma.

2. MATERIAL

- 1 Slide
- 1 Cover slip
- Distilled water
- 10% Salt water
- Scissors
- Needle

3. PROCEDURE

- PLANT CELLS OBSERVATION:

1. Cut the stalk of the leek.
2. In the place of the cut, pull out the transparent part of the epidermis using forceps.
3. Using the brush, place the peel onto the slide containing a drop of tap water.
4. Take a cover slip and place it gently on the peel with the aid of a needle.
5. View it in the microscope.
6. Describe the change in the shape of the cells.
7. Draw a diagram with the parts of a stome: stoma,cell guards,epidermis cells.

- SALT TREATMENT:

1. Prepare a 10% of salt solution.
2. Put the salt with a dropper on the left part of the slide.
3. Place a piece of cellulose paper in the opposite part of the cover slip, and let the dissolution to go though you sample.

4. RESULTS AND OBSERVATIONS

L15: ANIMAL CELLS vs PLANT CELLS

1. OBJECTIVES

- Identify the major components of cells.
- Differenciate between animal and plant cells,
- Measure dimensions of the entire cell and the nucleous.

2. MATERIALS

- Toothpick
- 2 Slides
- 2 Covers slips
- Distilled water
- Methylene blue
- Iodine
- Onion
- Glycerine
- Two whatch glasses
- Dropper
- Needle

3. PROCEDURE

- PLANT CELLS OBSERVATION:

1. Pour some distilled water into watch glass.
2. Peel off the leaf from half a piece of onion and using forceps, pull out a piece of transparent onion peel (epidermis) from the leaf.
3. Put the epidermis in the watch glass containing distilled water.
4. Take a few drops of iodine solution ( or safranin) in a dropper and transfer into another watch glass.
5. Using a brush ( or a needle), transfer the peel into the watch glass containing the dye, Let this remain in the safranin solution (or iodine) for 30 seconds, so that the peel is stained.
6. Take the peel from the iodine solution and place it in the watch glass containing distilled water.
7. Take a few drops of glycerine in a dropper and pour 2-3 drops at the center of a dry glass slide.
8. Using the brush, place the peel onto the slide containing glycerine.
9. Take a cover slip and place it gently on the peel with the aid of a needle.
10. Remove the extra glycerine using cellulose paper.
11. View it in microscope.

- CHEEK CELLS OBSERVATION:

1. Gently scrape the inner side of the cheek using a toothpick, wich will collect some cheek cells.
2. Place the cells on a glass slide that has water on it.
3. Mix the water and the cheek cells using a needle and spread them.
4. Dry the sample under the light to fix the sample on the slide,
5. Take a few drops of methylene blue solution using a dropper and add this to the mixture on the slide,
6. After 2-3 minutes remove any excess water and satin from the slide using cellulose paper.
7. Take a clean cover slip and lower it carefully on the mixture with the aid of a needle.
8. Using the top of the needle, press the cover slip gently to spread the epithetial cells,
9. Remove any extra liquid around the cover slip using cellulose paper.

4. RESULTS AND CONCLUSIONS

PLANT CELLS:

MRcell: 6,9/400= 0,017cm
0,17x10000= 172,5microns

MRnucleous; 400=0,7x10000/X
X=7000/400=17,5 microns


CHEEK CELLS:
400= 1,5x10000microns/X
X=1,5x10000microns/400=37,5

400=0,3x10000microns/X
X= 0,3x10000microns/400=7,5microns

L12: DNA EXTRACTION

1. INTRODUCTION

Deoxyribonucleic acid (DNA) is a nucleic acid that encodes the genetic instructions used in the development and functioning of all known living organisms and many viruses. Nucleic acid are biopolymers formed by simple units called nucleotides. Each nucleoide is composed of a nitrogen- containig nucleobase ( G,T,C,A) as well as a monosacharide and a phosphate group.
These nucleoides are joined to one another in a chain by covalent bonds between the sugar of one nucleoide and the phospate of the next. Most DNA molecules consist of two strands coiled around each other to form a doble helix. Hydrogen bonds bind the nitrogenous bases of the two separate strands.
The two strands run iin opposite directions to each other and are therefore anti-patallel. Moreover the bases of the two opposite strands unit according to base pairing: A-T and C-G.
Within cells, DNA is organized into structures called chromosomes.

2. OBJECTIVES

- Study DNA structures.
- Understand the process of extracting DNA from a tissue.

3. MATERIAL

- 1L Erlenmeyer flask
- 100mL beaker
- 10mL graduated cylinder
- Small funnel
- Glass stirring rod
- 10mL Pipet
- Knife
- Safety goggles
- Cheesecloth
- Kiwi
- Pineapple juice
- Distilled water
- 90% Ethanol ice-cold
- 7mL DNA buffer
- 50mL dish soap
- 15g NaCl
- 900mL tap water

4. PROCEDUCE

1. Peel the kiwi and chop it to small pieces. Place the pieces of the kiwi in one 600mL beaker and smash with a fork until it becomes a juice pure.

2. Add 8mL of buffer to the mortar.

3. Mash the kiwi puree carefully for 1 minute without creating many bubbles.

4. Filter the mixture: put the funnel on top of the graduated cylinder. Place the cheesecloth on top of the funnel.

5. Add beaker contain carefully on top of cheesecloth to fill the graduated cylinder. The juice will drain through the cheesecloth but the chucks of kiwi will not pass through in to the graduated cylinder.

6. Add the pineapple juice to the green juice ( you will need about 1mL of pineapple juice to 5mL of the green mixture DNA solution). This step will help us to obtain a purer solution of DNA . Pineapple juice contains an enzyme that breaks down the proteins.

7. Tilt the graduated cylinder and pour in an equal amount of ethanol with an automatic pipet. Put the ethanol through the sides of the graduated cylinder very carefully. You will need about equal volumes of DNA solution to ethanol.

 8. Place the graduated cylinder so that it is eye level. Using the stirring rod, collect DNA at the boundary of the ethanol and kiwi juice; only the stir in the above ethanol layer!

9. The DNA precipitate looks like long, white and thin fibers.

10. Gently remove the stirring rod and examine what the DNA looks like.

5. OBSERVATIONS/ CONCLUSION

We can observe the fibers of DNA.

6. QUESTIONS

1. What did the DNA looks like?

The DNA looks like white  and thin fibers.

2. Why do you mash the DNA? Where it is located inside the cells?

The crush to extract the liquid from the kiwi, the DNA is in the nucleus of cells.

3. DNA is soluble in water, but not in ethanol. What does this fact have to do with our method of extraction?

We can see the DNA in the part of ethanol because, if we touch the water the DNA can dissolve.

L11: PROTEIN AND EVOLUTION

1. INTRODUCTION

Genes are made of DNA and are inherited from parent to offspring. Some DNA sequences code for mRNA wich, in turn, codes for the amino acid squence of proteins, Cytochrome C is a protein involved in using energy in the cell. Cytochrome C is found in most, if not all, known eukaryotes. Over time, random mutacions in the DNA sequence occur. As a result, the amino acid sequence of Cytochrome C also changes. Cells without usuable C are unlikely to survive.

2. OBJECTIVES

To compare the reladness between organisms by examining the amino acid sequence in the protein, Cytochrom C.

3. PROCEDUCE

 The image is from Ignacio, because I can not find my photocopy.

4. CONCLUSION

As far from each other and are more differences further evolutionarily amino acids. Are grouped by groups of animals.

5. QUESTIONS

1. How many Cytochrom C amino acid sequence differences are there between chickens and tukeys?

0.

2. Make a branking tree, or cladrogram for chikens, penguins and turrrkeys?

chicken- penguin: 0
turkey-penguin: 3

3.a Predict the number of Cytochrom C amino acid sequence differences you would expect to see between.

Horse-Zebra: 1-2

Donkey-Zebra:1-2

b) What other information did you use to make this prediction?

If they can repoduce and the offpring is festile.

5. List three other things used to determine how organisms are related to each other.

Comparing the organs, anatomic prove, embrions,,,

6.  Explain why more closely related organisms have more similar Cytochrome C.

Evolutionarily not so long ago parted hence have not so many mutations.

7. Other data, including other genes, suggest that fugi are more closely related to animals than plants. What are some reasons that the Cytochrome C data suggest that fugi, plants, and animals are equally distantly related?

If you have more than 40, suffered a lot of mutations and can not be compared. 

L10: PROTEIN DESNATURALITION I

1. INTRODUCTION

Desnaturation is a process in which proteins or nucleic acids lose the quaternary, tertiary and secondary structure that is present in their native state. Desnaturation is the result of the application of some external stress ( heat and pH change) or base, a concentrated inorganic salt or organic solvent.

If proteins in a living cell are denatured, this results is disruption of cell activity and possibly cell death. Denatured proteins can exhibit a wide range of ring characteristics, from loss of solubility to communal aggregation.
This las effect results from the bonding of the hydrophobic proteins to reduce the total area exposed to water. In very few cases denaturation is reversible and proteins can recuperate their native state when the denaturation is reversible and proteins can recuperate their native state when the denaturing factor is removed. This process is called renaturation.

2. OBJECTIVES

- Study the relation between the structure and the function of proteins,

- Understand how temperature, pH snd salinity affect to the protein structure.

3. MATERIAL

- 2X50 mL beaker
- 4 test tubes
- Test tubes rack
- 10 mL pipet
- Knife
- Glass marking pen
- Potato
- Distilled water
- Hydrogen peroxide
- NaCl
- HCl

3. PROCEDUCE

In this experiment we are going to test the catalase activity in different environment situacions. We are going to measure the rate of enzyme activity under various conditions, such as different pH values and temperatures. We will mesuare catalase activity by observing the oxygen gas bubbles when H2O2 is destroyed. If lots of bubbles are produced, it means the reaction is happening and the catalase enzyme is very active,

-  Prepare 30 mL of H2O2 10% in a beaker ( use a pipet).
-  Prepare 20 mL of HCl 10% in a beaker.
-  Prepare 30 mL of NaCl 50 % in a beaker.
-  Peel a fresh potato tuber and put the tissue.
- Label 5 test tubes (1,2,3,4,5).
- Immerse 10 minutes your piece of potato inside HCl and NaOH beaker, and mashed up the potato.
- Add 5 mLH2O2 10% in each test tube.
- With a glass- marking pen mark the height of the height of the bubbles.
- Compare the results of the 5 test tubes.

4. OBSERVATIONS 

- Independent variable: treatment of each potato.
- Dependent variable: the height of the bubbles.
- Experimental Group(s): the rest.
- Control Groups: treatment of each potato.
- Constants: size, amount of H2O2, time...

5. CONCLUSION

Mashed > Raw > NaCl > HCl > Potato boiled.

6. QUESTIONS

1. How did the temperature of potato affect the activity of catalase?

The enzime X of the catalase.

2. How did the change of the Ph of the potato affect the activity of the catalase?

The pH change for acid the catalase no activity while ppH change for a basic the catalase activity.

3. In wich potato treatment was catalase the most active? Why do you think this was?

The mashed up potato , because when we mashed up the potato break the enzyme X.

L9: PROTEIN IDENTIFICATION

1. INTRODUCTION

Biuret's test is a chemical test used for detecting the presence of peptide bonds. A peptide bond can be broken by hydrolysis (the adding of water). In organisms, protein molecules called enzymes facilate the process.

The biuret reaction can be used to assess the concentracion of proteins because peptide bonds occur with the same frequency of amino acid peptide. 

The solution to be tested in treated with a strong base followed by a few with a strong based followed by a few drops of copper (II) sulphate. If the solution turns purple, protein is present.

2. OBJECTIVES

- Identify peptide bonds.

- Compare protein concentracion in differents foods.

3.MATERIAL

- 7 x 250mL beaker

- 6 test tubes

- Test tube rack

- 6 x 10mL Pipet

- Mortar

- Glass marking pen

- Gloves

- Goggles

- Milk

- Soy milk

- Egg

- Yogurt 

- Potato

- Distilled water

- NaOH 20%

- 10 drops of CuSO4

4. PROCEDURE

First of all are going to dilute the protein,

1. Add 100mL of distilled water to each 250mL beaker. Label them with M (milk), S (soy milk) and EW ( egg white), EY (yolk), Y (yogurt) and P (potato).

2. Separate the egg white nd the yolk in another beaker.

3. Smash the potato in a mortar and add some amount of the smashed potato to the P beaker.

Prepare the samples:

4. Add 10 mL of a dispersion of each food ( M,S,EY,EW and Y) to the indicate beaker. Calculate the final concentracion. All the groups will use the same dispersion from the beakers.

5. Prepare 6 test tubes and label M,S,EW,EY,Y and P. Add 2 mL to test tube of the every food disolution of each beaker.

6. Add 2 mL of 20% NaOH dissolution.

7. Shake gently and add 5 drops of CuCO4 in each tube. Allow the mixture to stand for 5 minutes.

8. Note any colour change. Remember that proteins will turn solution pink or purple.

9. Compare the test tubes.

5. OBSERVATIONS

6. QUESTIONS

1. Which food has protein? 

Animal food.

2. Which food has more proteins? Why?

Eggs white, yogurt, potato and milk. Are animal food.

3. Do you find any different between soy and cow milk?

Rice milk, doesn't have protein, however has milk has.

4. Is there any different among milk and yogurt?Why?

Yes. Yogurt has more protein than milk.