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

These notes are provided to assist in the preparation
and execution of the laboratory experiment.
A solutions key for the pre- and post-lab
questions can be found on the reverse.
Supplies
First, inventory the supplies included in the lab kit.
Supplies have been provided for up to six groups of
students.
• DNA for Known Gels (3 sets)
• DNA for Evidence Gels (3 sets)
• Agarose Gels (6 bottles)
• 20x BlueVis Buffer (2 bottles)
• BlueVis Gel Stain (1 bottle)
• InstaStain Sheets (6 sheets)
Refrigerate the entire contents of the kit until ready
to use.
Other Supplies and Equipment Required
• Lab gloves
• Distilled water
• 1 to 4 L capacity bottle
• Microwave
• 5 mL Pipettes and pipettor
• Gel apparatus (6) and power supply
• 10 μL Pipettor and Pipette tips
• OPTIONAL: Staining trays
Running Buffer Preparation
The 20x BlueVis Buffer running buffer must be diluted
to 1x before being used in the lab. Each 200
mL bottle of buffer will make 4 L of the solution
used in this lab.
Shake the Blue Vis Buffer well before use. To make
1 L of 1x BlueVis Buffer add 50 mL of the 20x stock
solution to a 1 L capacity bottle, and bring the total
volume up to 1 L with distilled water.
Safety Precautions
1.
2.
3.
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5.
Teacher’s Notes
Molten agarose can scald and it must be handled
with care
Serious or lethal electrical shock can result if
the gel apparatus is used improperly.
Remove caps from bottles placed in the microwave
to avoid explosion and serious injury.
BlueVis buffer and Gel Stain will stain skin and
clothing; wear gloves while handling.
Review MSDS’s found on the Mystery of Lyle
and Louise website.
http://www.lyleandlouise.com/msds
Pipetting Practice
Inexperienced students should practice pipetting
before performing this lab. One suggestion is to use
a small piece of waxed paper and practice picking
up and delivering small volumes of colored water.
Students should hold the pipettor as vertical as
possible. Without precise pipetting, experimental
results may not be accurate.
Notes for the Gel
The kit includes enough samples for 3 groups to
analyze known suspects and victims (Gel A) and another
3 groups to analyze the unknowns and evidence
(Gel B). DNA labels are color-coded to keep
them separate in the lab; DNA that should be run on
Gel A are in blue, DNA for Gel B are in red. Ladder
DNA, which must be run on both gels, has a white
cap.
The BlueVis Gel Stain is the same solution as the
20x BlueVis TBE running buffer, but is bottled separately
to prevent confusion. If more of the stain is
needed, remaining 20X BlueVis Buffer can be used
with no problem. Gels thicker than 1 cm have poor
contrast. Use less than the 80 mL of agarose provided
if you have small gel boxes. Note: BlueVis will
not stain DNA if it is overheated. Don’t remelt agarose
after adding BlueVis.
The Mystery of Lyle and Louise 7