Bio 243: LABS

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Laboratory One: What are genes made of?

CONSTRUCTING DNA:

Construct 4 types of deoxynucleotides

Connect A and T deoxynucleotides together

Connect the next four nucleotides

Build a hexanucleotide with a complementary sequence

Make the strands antiparallel to one another

Construct base pairs and twist to make helical shape

EXTRACTING DNA:

Crush strawberry

Add the DNA extraction buffer

Set up a funnel and cheesecloth and pour in mixture, let filter

Label centrifuge tubes and add in respective materials

Pour isopropanol into original tube

Stir the mixture and withdraw at intervals

Dip pipette into tube A and clean carefully

Dip pipette into tube B and stir, then transfer solution to original

Add dye to all three small tubes and add DNA to two of them

Spot mixture from all three small tubes

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Laboratory Two: Genetic Engineering and UV Radiation

TRANSFORMING E. COLI:

Wipe down and collect materials

Label the microcentrifuge tubes and put them in the rack

Pipette transforming solution

Put all tubes over ice

Sterilize loop and pick up bacteria

Vortex and place on ice

Repeat for the other sample

Add aliquot, close and vortex, put on ice

Incubate on ice

Label agar plates

Heat shock organisms

Add broth on bench

Close and vortex, incubate

Gently vortex, pipette both samples on plates

Sterilize spreader and add alcohol, burn

Spread on plates, sterilize, cool

Incubate plates when dry

Wipe down and wash hands

EXPOSE YEAST:

Wipe down and label plates

Vortex and pipette, spread

Repeat for each plate

Sterilize and let cool

Prepare templates and collect materials

Tape and label templates, use foil

Weigh both sunscreens and spread

Remove lids and place in UV hood

Clean up materials and dispose of containers

Wipe down and wash hands

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Laboratory Three: Mutations

EFFECTS OF UV RADIATION:

Obtain incubated plates

Count the yeast colonies

Record data

Return plates

MUTATIONS:

TA to choose number

Roll dice and write in row

Fill out results column

Repeat steps 1-3 up to seven more times

Tally the results

Record the pooled data

GENETIC ENGINEERING:

Obtain incubated plates

Count the yeast colonies

Record data

Return plates

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Laboratory Four: Process of Mitosis

STAGES OF MITOSIS

Place slide under microscope and centre the field of view

Focus on root tip and add a drop of oil

Focus on higher magnification

Clean slides

EFFECTS OF CAFFEINE ON MITOSIS

Cut the control root tip and put into labelled microcentrifuge tube

Cut three root tips and put into labelled microcentrifuge tube

Add HCl to each tube, close, and incubate

Place four slides on paper towel and label

Remove root and place on slide

Add two drops of water, then stain, then water, then acid, then water

Place coverslip on root

Use microscope to look at root tip

Place drop of oil, choose cells and count

Clean up and dispose of materials

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Laboratory Five: Meiosis

SALINITY GRADIENT

Wipe done bench and wash hands

Obtain dish and label with arrows and names

Obtain the saline-nutrient agar

Pour and close lid

Swirl petri dish and raise one end

Leave for 0 minutes

Obtain the nutrient agar and mix it

Pour onto high part of agar dish

Leave for 25 minutes

STAGES OF MEIOSIS

Assemble diploid model cell with materials

Place sticker alleles on model

Place adhesive on centromere

Attach second copy of models

Pair up homologous chromosomes

Move to different sides of metaphase plate

Line up centromere with tape

Select gametes and combine

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Laboratory Six: Evolution in Populations

INNOCULATING SALINITY AGAR PLATES

Obtain the plate used last lab and a control plate

Label the control plate and draw an arrow

Turn both of the petri dishes the right way

Place a ruler underneath

Obtain and vortex the bacteria

Pipette 6 droplets 1.5cm apart on the plate

Replace lid and set plate aside

Vortex nutrient broth

Stack both plates and place them in container 1

When dry, place in container 2

Wipe bench and wash hands

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Laboratory Seven: Statistics and Hypotheses

PREPARING S3 PLATE

Prepare plate in the same way as procedure 5

ANALYZING S1 PLATES

Obtain the S1 and C1 plates and turn them the same way

Measure the distance from the edge to the last colony with a ruler

Mark the bottom of the plate with the location

Determine a conversion factor

Create a table to record the data

INNOCULATE S2 PLATES

Use a cooled sterile tip to pick up sample

Lift S2 lid an drag the loop across the surface

Wrap parafilm around the S1 plates

Stack the plates and put them away

Wipe bench and wash hands

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Laboratory Eight: Evolution in Bacteria 3

ANALYZING S2 PLATES

Wipe down bench

Obtain plates

Record distance from edge of plate to last colony

Convert distance to saline concentration

Record data

INOCULATING S3 PLATES

Obtain plate and control plate

Label control plate and draw an arrow

Streak cells from colonies in high salinity to low salinity

Wrap parafilm around plates

Stack plates and put them away

Wipe down bench and wash hands