 
Electrophoresis is the process of separating large 
macromolecules, typically DNA, RNA, and proteins. 
It was originally developed in the 1940s employing a 
roomful of equipment 
and popularized in the 1950s by Oliver Smithies. 
Today, it has been refined to a small desktop unit 
but it is still somewhat expensive. 
While high school students throughout North Carolina 
are expected to learn this process, 
very few schools can afford a gel electrophoresis kit. 
 DIY biology is a growing movement in which individuals 
study biology and life science using the same methods as 
traditional research institutions. 
These citizen scientists get involved as a hobby 
or as a non-profit endeavor for community learning 
and open-science innovation. 
 Recently, the National Institute of Environmental Health Sciences 
hosted workshops and seminars for science teachers 
in North Carolina. 
As part of the workshop, Triangle DIY Biology presented 
a working session, showing a method of creating 
an electrophoresis experiment using equipment and resources 
that are readily available and affordable. 
 In this video, we show the details of that working session,
demonstrating how to build and run a basic 
gel electrophoresis experiment using items that you 
can find around the house or purchase from any grocery store. 
Agar for the gel can be found in most Asian food markets. 
 For our kit, we will be conducting food color electrophoresis
 to look at the separation of food color  dyes. 
Colored food dyes are made of small chemicals of different 
sizes and different charges and like proteins and DNA,
 they migrate through the agar gel when placed in 
an electrical current at different rates. 
 The first step is to make the gel in which our dye will migrate. 
 In a flask, we mix 100 milliliters water and 
0.2 grams of baking soda. 
Just swirl the mixture by hand.
Then we add 1 gram of agar and swirl again until it is mixed. 
 Place in microwave and heat for about a minute 
checking carefully that it does not boil over. 
Keep checking and return to the microwave until 
the solution is clear and not granular in any way. 
Let cool for 15-20 minutes. 
 After cooling, we're ready to make the mold. 
Our mold is made from the lid of a pipette tip box. 
The gel comb is made using a laser cutter or a 3-D printer. 
Pre-made combs are available online from many 
DIY biology groups including Triangle DIY Biology. 
 Place the comb in the mold and slowly pour 
all the mixture into the mold. 
Let this set for 1 - 2 hours. 
 While we're waiting for the mold to set 
we can prepare the dyes. 
For this we use water, corn syrup and food dyes 
in a culture plate. 
 Put some water into a beaker or any other container 
and place 3 milliliters of water in six of the reservoirs.
 Next add 0.5 milliliters of corn syrup for each. 
This adds density to the mixture. 
 Lastly, add 2 drops of food dye. 
You can experiment with different combinations of colors, 
but here we're using 4 single color and 2 mixed colors. 
 For some students it also fun to use candy 
such as Skittles or M&Ms instead of dye. 
Kids enjoy seeing what their candy is really made of! 
 Now our mold should be completely cool and firm to the touch. 
Very gently remove the comb. 
This creates six wells where we will load our sample mixture
 we just made while we were waiting for the mold to cool. 
 Using a sharp knife or blade cut the ends of the mold to 
make room for the electrodes from your power supply. 
Use the spatula to remove the excess gel and discard. 
Keep in mind that there is nothing toxic about any of these substances. 
 Now fill the mold with a buffer solution of 
.2 grams baking soda to 100ml of water or .2%. 
Make sure the gel and the wells are entirely submerged. 
 Crimp 1 millimeter stainless steel wires so they fit 
into each channel we just created in the gel 
with the ends facing up. 
Paper clips are acceptable in a pinch but they corrode very 
quickly and stain the mold so stainless steel is preferred. 
 The power source is a series of five standard 
9-volt batteries daisy-chained together. 
Keep in mind the voltage here is extremely low and 
there is a minimal risk of shock. 
 Connect the positive end of the batteries to the wire 
FARTHEST from the wells with the alligator clips. 
Connect the negative end of the batteries to the wire 
CLOSEST to the wells. 
The dye will migrate from the negative charge 
to the positive charge. 
DO NOT connect the negative end to the battery yet! 
This will be done after we load the dyes into the wells. 
 Draw a few drops of your dye mixture into a pipette 
and load it into a well. 
Place the pipette into the well before releasing the dye. 
It's ok to overload a little since this will diffuse away 
during the run and won't affect the result. 
Load each of the six wells with one of the six 
dye mixtures that was made. 
As soon as the samples are loaded, complete the circuit 
to begin the run. 
 Once the process begins and power is flowing through 
the mold you will see bubbles coming off of the wire.
That's how you know it's working! 
Let run for one and a half hours. 
The dyes have a negative charge so they migrate 
to the positive end over time. 
The molecules of the different color dyes are different 
molecular weights; the smallest molecules moving faster 
through the dye than the larger ones. 
You can see how this separates the various colors from each well. 
 And that's how you can make an inexpensive agar gel 
electrophoresis kit using everyday items you can easily locate. 
We hope you have enjoyed this video and 
good luck with your electrophoresis! 
