Link to our lab as a pdf - College of Science - Marshall University

Lab Procedure
Preparing the Gel (45 min)
1.
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8.
Remove the cap from the bottle of agarose gel
and microwave in 15-30 second intervals until
the agarose has completely melted. Watch that
it does not over-boil in the bottle. Let the agarose
cool to at least 60°C before continuing.
Shake the bottle of BlueVis Gel Stain well.
Transfer 4 mL of BlueVis Gel Stain into the bottle
with the agarose. Recap and swirl to incorporate
the stain thoroughly.
Position the gel box on a level surface and insert
the gel tray, ensuring that all sides are
tightly sealed.
Place the comb at one end of the gel tray.
Slowly pour the agarose into the gel tray until
the central cavity and the crevices between
each tooth in the comb are filled.
Wait until the gel has become solid and slightly
opaque (approximately 20—30 minutes).
Remove and turn the gel tray so that the wells
are near the black (-) electrode and the rest of
the gel points to the red (+) electrode.
Pipetting Order
Gel A
Ladder
Lyle Mondelo
Louise Mondelo
Wally Mondelo
Jan Mondelo
John Wayne Gretzky
Preparing the Samples
Gel B
Ladder
Adult Woman (Car)
Female Child (Car)
Male Child (Car)
Adult Woman (Cabin)
Adult Male (Cabin)
Firewood (Cabin)
Fireplace (Cabin)
The samples will be divided over two gels, A and B,
to keep the knowns separate from the unknowns/
evidence. Both Gels A and B will be run in triplicate;
that is, three groups will run a copy of gel A
and three groups will run a copy of gel B.
Gel A will contain samples that are known; DNA
for these samples is labeled in Blue. Gel B contains
unknown samples from the cabin and car accident;
these samples are labeled in Red. The ladder has
five lengths of DNA at 150, 300, 500, 650, and 1000
base pairs. It is labeled in Black and will be run in
the first lane of both gels.
1.
2.
Collect a set of samples for the gel you will be
running and set them before you.
Order your samples according to their pipetting
order.
9.
Fill the gel box with enough 1x BlueVis buffer
solution provided by your teacher to completely
cover the gel, making sure that the wells are
filled.
10. Carefully remove the comb.
Loading the Gel
1. Place a new tip on the pipettor. This should be
done before each sample.
2. Ensure pipettor is set to 10 μL.
3. Tap the sample tube against the palm of your
hand to settle the contents at the bottom.
4. Open the sample tube.
5. Press down on the pipette plunger until you
reach the first stop. The plunger can continue
The Mystery of Lyle and Louise 27