BIO 100 LAB PREP
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KINGDOM MONERA LAB
KINGDOM MONERA - WEEK TWO
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KINGDOM MONERA LAB

MATERIALS FOR LAB:
 
6     200 ML BEAKERS
6     100 ML GRADUATED CYLINDER
6     CELCIUS THERMOMETERS
6     MEASURING TEASPOON
6     MEASURING TABLESPOON
6     PLATES (LOCATED ON EACH TABLE IN THE ROOM)
24    BOXES OF MICROSCOPE SLIDES (IN DRAWERS)
24    BOXES OF COVERSLIPS (IN DRAWERS)
6      PAPER CUPS (1/2 TO 1 CUP SIZE)
6      BOTTLES OF METHYLENE BLUE STAIN
6      TONGUE DEPRESSORS
1      ROLL OF SARAN WRAP
6      PH PAPER (WIDE RANGE NOT LITMUS)
6      RUBBER BANDS
1      SMALL ROOM TEMPERATURE INCUBATOR
6      1 LITER PLASTIC POP BOTTLES WITH LIDS
5      HEADS OF RED CABBAGE CUT UP INTO SMALL PIECES
1      SHARP BUTCHER KNIFE
1      MEDIUM TO LARGE BAG OF COARSE SALT (NOT TABLE)
6      MORTAR AND PESTLE
6      3 INCH TEST TUBE
6      ROLLS OF 1/2 INCH ADHESIVE TAPE
6      PIECES OF 12 INCH COPPER WIRE
1      WIRE CUTTER
1      ROLL OF COPPER WIRE
1      AQUARIUM FULL OF WATER SEEDED WITH BACILLUS MEGATERIUM (PLACED IN THE BACK OF THE ROOM)
24     MICROSCOPES
 
1 BOX OF POWDERED MILK
1 CONTAINER OF YOGURT WITH ACTIVE CULTURES PER LAB PERIOD
YOU ALSO NEED TO MAKE SURE THAT THE PAPER TOWEL RACKS ARE FILLED WITH PAPER.  IF NOT THEN PLACE A STACK OF PAPER TOWELS AT THE FRONT OF THE ROOM.

BACTERIAL FERMENTATION—YOGURT

 

LAB PERIOD ONE    13 OCT 03 (This is a Monday Lab)  

 

Each group will need

 

1          200 ml beaker

1          100 ml graduated cylinder

1             Celcius thermometer

1          teaspoon

1            tongue depressor (for stirring)

            powdered milk

            yogurt culture

            hot plate

           

Examine the prepared yogurt culture provided to you.  Note its color, aroma and texture. Make a wet mount of a small amount of the culture, using Methylene Blue as your stain. 

 

 

 

 

 

 

 

 

 

 

 

 

  DRAW WHAT YOU SEE HERE

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 


COLOR    ____________________________________________________________________

              

   ____________________________________________________________________

 

               ____________________________________________________________________

 

 

AROMA   ____________________________________________________________________

 

               ____________________________________________________________________

 

               ____________________________________________________________________

 

TEXTURE ___________________________________________________________________

 

               ____________________________________________________________________

 

               ____________________________________________________________________

 

PROCEDURE

 

Add 100 ml of water into a 250 ml beaker and heat until it boils. As the water is cooling,  measure out 7 tablespoons of powdered milk.  When the water cools to 45 degrees C, add the powdered milk and stir thoroughly with the tongue depressor.  Next add one teaspoon of yogurt culture and again stir thoroughly.  Transfer the culture to a small paper plate, cover with saran wrap and secure it in place with a rubber band and incubate at room temperature.

   BACTERIAL FERMENTATION—YOGURT

 

LAB PERIOD TWO  (OCT 20)

 

Examine your yogurt culture.  Note its color, aroma, and texture.  Can you explain why these may be different from the commercially prepared cultures.  Make a wet mount of a small amount of the culture, using methylene blue as your stain.  Compare the drawing of your commercially prepared yogurt with your drawing of your culture.  Are they the same or different?__

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

DRAW WHAT YOU SEE

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 


COLOR    ____________________________________________________________________

               ____________________________________________________________________

               ____________________________________________________________________

 

AROMA  _ ___________________________________________________________________

               ____________________________________________________________________

               ____________________________________________________________________

 

 

TEXTURE ___________________________________________________________________

               ____________________________________________________________________

               ____________________________________________________________________

BACTERIAL FERMENTATION--SAURKRAUT

OCT 13(This is a Monday Lab)

 

Saurkraut is a favorite food dish in Korea.  It is an example of pickling foods.  The process occurs due to the action of bacteria that produce lactic acid during the process of fermentation.  Sugars in the cabbage leaves is converted to lactic acid.  This lowers the pH and produces an acidic environment that prevents the growth of food spoiling organisms. 

 

EACH TEAM OF STUDENTS WILL NEED

 

1          Saurkraut container  (1 liter bottle)

½         head of cabbage

25 ml of salt

1          mortar

1          3 inch test tube

            pH indicator paper (wide range)

 

PROCEDURE:

 

1.                  Assemble all the necessary equipment and supplies

 

2.                  Place a layer of cabbage in the bottom of the Saurkraut container

 

3.                  Sprinkle a small amount of salt on the layer of cabbage

 

4.                  Use the mortar to tamp down the layers.

 

5.                  Continue to fill the container until it is 2/3 full, alternating the cabbage with salt and tamping down each layer. 

 

6.                  Set the mixture aside and continue to tamp down the contents every 15 minutes for the next hour. 

 

7.                  Check the pH of the mixture.

 

8.                  Add the bottle top to the Saurkraut container.  Be to slide the top down  until the cabbage juice rises above the Petri dish.  This will seal the container and allow the fermentation process to occur.

 

9.                  Check the progress of your culture each day.  Be sure to press on the top of the container to keep the cabbage covered with fluid.

 

10.              Measure the pH of the medium each day.

 

11.              Incubate for 7 days

 

 

 

 

 

SAURKRAUT OBSERVATIONS

Oct 20

 

DAY

pH

Description

 

 

1

 

 

 

 

 

 

 

 

 

2

 

 

 

 

 

 

 

 

 

3

 

 

 

 

 

 

 

 

 

4

 

 

 

 

 

 

 

 

 

5

 

 

 

 

 

 

 

 

 

6

 

 

 

 

 

 

 

 

 

7

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

BACTERIAL FERMENTATION--SAURKRAUT

 
LAB PERIOD TWO  OCT 20

 

 

Examine your Saurkraut.  Note its color, aroma, and texture.  Can you explain why these are different from the cabbage you originally started with?  Has the liquid portion changed colors?  If it has, can you explain why. 

 

Make a wet mount of a small amount of the liquid, using methylene blue as your stain.  Draw what you see below.

 

 

 

 



FISHING FOR BACTERIA

 

LAB PERIOD ONE   13 OCT (This is a Monday Lab)

 

Each group will need:                                                   

 

½ inch adhesive tape

2 microscope slides

1 copper wire (12nches long)

 

 

PROCEDURES

 

Taking your 12inch piece of copper wire, form  one end into a hook.  Tape the copper wire to the side of one of the microscope slides.  Be careful not to touch the flat surfaces of the slide.  Place the other slide on to the back of the first slide and tape all four edges together.  Write your group name on another piece of adhesive tape and attach it to your wire.

 

Hang your slide over the edge of the aquarium in the back of the room and let it sit until the next lab period.

 

 

LAB PERIOD TWO 20 OCT

 

Extract your slides from the aquarium.  Carefully, remove the adhesive tape and lay the slides aside to dry.  Be sure to have the exposed surfaces of the slide up.  After the slides have dried, prepare a gram stain and examine the various types of bacteria present on the slide.  Draw what you see below.

 

 

 

 

 



LAB PROCEDURE -- PREPARING A GRAM STAIN

 

 You will prepare a bacterial smear.  Before beginning be sure to:

 

      1.   Prepare your slide by passing them through a Bunsen three times.

 

  1. Allow it to cool.

 

  1. Cover the smear with crystal violet stain for one minute.

 

  1. Quickly wash off the stain using the bottle of distilled water.  Shake off excess water.

 

  1. Cover the smear with Gram’s iodine for one minute.

 

  1. Shake off the Gram’s iodine.

 

  1. Rinse  the smear with 95% Ethanol until the runoff is clear.

 

  1. Rinse the Ethanol off with water.

 

  1. Cover the smear with safranin for one minute.

 

  1. Rinse the smear with water for 10 seconds.

 

  1. Blot the smear with paper toweling and let it air dry. 

 

  1. Draw what you see under 40x

 

Drawings will be due at end of lab

20 OCT.
MATERIALS NEEDED FROM THE STOCKROOM FOR OCT 20

 

GRAM STAINING KITS

 

WE WILL BE FINISHING THE LAB FROM THE PREVIOUS WEEK

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