TRANSFORMATION (13 min)

Poster – Describe transformation

            Bacteria (E. coli, in your gut)

            Plasmid – little extra piece of DNA in a bacteria cell

Describe Ampicillan (antibiotic….kills bacteria)

Resistance vs. Susceptible

Plasmid

-         this one = pGLO

-         has a portion of a gene from a glowing jellyfish

-         Makes E. coli resistant to amp.

When arabinose present = portion of the plasmid with jellyfish gene turned on and bacteria glow – NEED UV LIGHT to make bacteria glow

WHY TRANSFORMATION GOOD!

            Bacteria can make human form of insulin

make foods more nutritious (not a problem in this country, but very important for poorer

countries)

Have students make slides

Let’s look at different kinds of cells and YOUR cells

            Plant cells

                        Elodea Leaf

            Algae cells (pond water sample)

            CHEEK CELLS

                        Little water – use toothpick to get cells from cheek and swirl in water and

then a small drop of Methylene Blue and then cover slip

 CANCER (13 min)

What makes a cancerous cell (keeps dividing, show poster of mitosis…why cells usually go through mitosis…growth and repair)

Why caner is bad (don’t scare themJ)

Cancer can be GOOD!

            Culture (raise) cancer cells – lines have been alive since 1950s!

Research

cell biology (how all the processes work)

            cancer

To keep cancer cells alive we must feed them

            PROCEDURES:

1.      Wash hands

2.      take media out of water bath and Pour off “top” app. 5ml

3.      take cells out of water bath

4.      Wipe of everything with Ethanol

5.      pipette 5ml of 37°C media onto the side wall of the flask

6.      Wipe off everything again and place cells back into incubator

Electrophoresis (13 min)

What electrophoresis is used for – looking at similarities in DNA

-         Each individual’s is unique – like a fingerprint

-         Criminal investigations

-         Relatedness among organisms

-         Human Genome Project

How Electrophoresis Works

-         DNA is cut into chunks

-         A sample is placed into a gel

-         The gel is submerged in a Buffer solution (keeps DNA together)

-         An electrical current is sent through the gel (DNA is negatively charged, it is attracted to the opposite positive side…runs toward red!)

-         Larger chunks “fall out” first

-         Show them pictures

How to do Electrophoresis

-         Get a sample of DNA (these came from Genetics students)

-         Amplify (make lots of copies using PCR)

-         Make gels (from agarose- a sugar that will form into this jelly)

-         Gel has wells (holes)

-         Mix blue juice with DNA (increases weight so it will fall down into holes)

-         Load DNA into wells

-         That what these guys are going to do 

o       8µl of DNA from PCR tubes

o       Put tip over well and slowly release liquid

o       Keep plunger down until lift out of water

o       Rinse out tip in water

-         Also, look at results! (TV magic = one already run for you)

***While waiting, students can practice micropipetting 8µl from one microcentrifuge tube into another

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Maintained by Linda Allen

3/20/07