Gel Electrophoresis

Using Food Dyes

Safety

ALWAYS wear your Goggles, when handling hot glass wear use
your GLOVES

DO NOT leave hot glass unattended

DO NOT plug in electrophoresis equipment

DO NOT open electrophoresis equipment

NO FOOD OR BEVERAGE

Exercise caution when using the power supply. The area

around the supply and the electrophoresis chamber should be
dry. Be sure all the connections are in place before turning on
the power. Likewise, the power supply should be shut off
before disconnecting any of the electrical leads.

Food dyes have an intrinsic SIZE and CHARGE

and thus can be separated using Electrophoresis
Food dye

Molecular
weight

Charge

FD&C Blue 1

792.86

-2

FD&C Yellow 5

534.37

-3

FD&C Yellow 6

452.37

-2

FD&C Red 40

496.43

-2

Carminic Acid

492.38

Beetroot red

551.48

+1

Lab Introduction

Agarose gel electrophoresis can resolve molecules

based on charge, size, and shape. In this laboratory
you will use gel electrophoresis to separate
molecules present in different food color mixtures.

FDA regulates color additives and approves only a

few different colors, from the research conducted
day one what are the colors recognized ?

Materials

Micropipetes and tips to load dye samples.

Dyes : Green, Blue, Red, Yellow, Unknown X, Unknown Z

Beaker ( for pipette disposal)

PPE (Goggles/gloves)

Hot plate

Erlenmeyer flask

Electrophoresis units and power supplies

0.8% Agarose in 1X TAE (melted)

1X TAE for electrophoresis units

Graduated Cylinder

Preparing the Gel

1.

Put the black rubber dams on the sides of the the gel deck. Be
careful to work them on slowly so you dont damage the deck.

2.

The gel will be melted on a hot plate. Careful it will be very hotuse gloves! Allow to sit on table until cool to touch.

3.

Carefully pour heated agarose into gel deck until the gel comes
close to the top. Do not overfill. Add comb.

4.

Put the deck out of the way and wait for the agarose gel to
solidify. Do not move it until it has hardened.

5.

Carefully remove black casting dams and comb.

6.

If the electrophoresis unit does not already contain buffer, pour

buffer in both wells and so that it covers the center section.

7.

Place the gel deck containing the gel in the electrophoresis unit.
Add buffer carefully so that the fluid level slightly covers the gel

1.0 % Agarose gel

Make a 1.0% Gel with 50 mL buffer

1x TAE buffer

Make a 1x Buffer by using a stock solution

The stock solution is

You want to make 500 mL

Pipetting the Sample

1.

Depress the operating button to the first stop.

2.

Immerse the tip slightly and release the operating button slowly. The tip
should be filled.

3.

Deliver the liquid by gently depressing the operating button to the first
stop. Then after a delay of about a second, continue to depress the
operating button all the way down to the second stop.

4.

Withdraw the tip and dispose in beaker

5.

DO NOT LAY THE MICROPIPETTE FLAT WHEN IT HAS A TIP ON IT!

THIS COULD CONTAMINATE THE PIPETTE

6.

Fill the pipet with 10 L of a food color sample. Place the tip over the top
of one of the wells. The tip should be submerged in the buffer at this
point. Holding the pipet steady, gently dispense the sample into the
well. The glycerin in the sample will allow the sample to sink into the
well. Do not place the pipet tip directly into the well or you will risk
poking a hole in the side or bottom of the well, and your sample may
leak out of the gel. A new pipet tip should be used for each sample.

Loading the samples

1.

Load the wells with the dye samples (R,Y, B, G) and the
unknowns (X and Z).

2.

Record which sample is in which well on your lab sheet.

3.

Carefully add buffer until it covers the gel.

4.

Make sure the power source is unplugged.

5.

Attach the cables from the chamber to the power source (red
to red and black to black).

6.

The instructor will adjust settings and turn on the machine.

7.

Run for 30-40 minutes.

8.

Unplug, take off the lid, and take out the gel bed containing
the gel and place it on a paper towel.

9.

Place the gel in the trash and leave the buffer in the chamber.*

Lab report
1. What is the purpose of gel electrophoresis?
2. What do you think are some factors determine the distance
molecules move though the gel?
3. In DNA fingerprinting, gel electrophoresis is used. A sample of a
persons DNA is cut at specific sites using substances called restriction
enzymes. The resulting fragments are of different lengths that are
characteristic of the individual the DNA came from. What is the role
of the gel electrophoresis in DNA fingerprinting?
4. Toward which poles (+ or -) on the electrophoresis apparatus did
the dyes move?
5. What charge do the dyes have?
6. What color dyes (different pigment molecules) make up each of the
food colors you used?
7. What were the unknowns?
8. What color pigment moved the farthest through the gel and why
did it go the farthest?
9. What pigment showed the least movement?
10. What are some practical uses of gel electrophoresis?