Gel Electrophoresis
Teacher Instructions
BioRad
Set Up
8 groups
Grossmont Kit
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Timeline
• Prepare Gels: Up to a week in advance
• Class lab: 1-3 days
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Teach students to pipette
Load and run gels
Teach electrophoresis theory
Analyze gels
• Gels must be analyzed no later than next day after
running (stored in refrigerator overnight)
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Prepare 1X TAE Buffer
for making gels
• Measure 28ml of 50X
TAE Buffer stock
solution into the 50ml
conical TAE Buffer
measuring tube
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Prepare 1X TAE Buffer
for making gels
• Pour the 28ml of 50X
TAE Buffer stock
solution into the 2L
TAE Buffer mixing
bottle
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Prepare 1X TAE Buffer
for making gels
• Fill 2L TAE Buffer
mixing bottle to the
1400ml line with water
(tap or distilled)
• You might need to repeat
this as necessary for your
number of classes – this
bottle should prepare
enough 1X TAE Buffer
for 6 classes worth of gels
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Prepare Agar for Gels
• Measure 3 level
scoops of agar powder
into each glass agar
mixing bottle
– Each scoop is 1/4 tsp
– This will equal ~1.5g
of powder per bottle
• You’ll need to make
1 bottle per class
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Prepare Agar for Gels
• Fill each bottle to the
200ml mark with your
prepared 1X TAE
Buffer solution
– Bottles have been prechecked for calibration
• Cap tightly and shake
to mix
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Prepare Agar for Gels
• Loosen caps slightly and
place bottles in your
microwave
• Set microwave for 1-2
minutes per bottle (less is
better - you can always
add more time!)
• Allow agar solution to
come to a boil - stop
microwave once a good
boil starts
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Prepare gels
• Carefully remove the
HOT bottle from the
microwave and swirl - be
careful of steam escaping
from the loose caps!
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Prepare Agar for Gels
• Check that agar has fully
dissolved or, if re-melting
solidified agar, that it has
all melted back into
solution
• Allow Agar to cool until
you can just stand holding
the bottle with your bare
hand
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Prepare Gel Casting Trays
• While Agar is cooling
use blue lab tape to
carefully seal the end
of each tray
• Place 2 combs into
slots on each tray (top
and middle)
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Pour Gels
• Carefully pour warm agar
solution into the
assembled gel casting
trays (make sure agar is
still fully melted)
• Fill each tray
approximately .75cm
• 1 bottle should fill 4 trays
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How to store prepared gels
• After gels have solidified,
remove the combs and
tape
• Carefully slide gel out of
the tray onto a “patty pac”
paper
• 1 gel per paper
• Re-tape tray to pour for
next class set
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How to store prepared gels
• Place 2 papers with 1 gel
each side by side in a gel
storage container
• Make sure paper edges are
free
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How to store prepared gels
• Stack second layer of gels
in storage container and
place container in fridge
for up to a week
• 1 container per class!
– (4 full size gels)
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Setting up prepared gels for class
• When you are ready to have
students use gels simply
carefully lift paper with gels
out of the storage container
• Carefully use spatula to cut
each gel in half (~1cm above
middle wells)
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Setting up prepared gels for class
• Place each gel half onto flat
tray for each student group
– Put a patty pac under if too hard
to see wells
• Try to keep gels and trays low
and level to prevent accidental
tearing of the gel
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Running gels
• Prepare 1X TAE Buffer
solution as before:
– Pour the 28ml of 50X TAE
Buffer stock solution into
the 2L TAE Buffer mixing
bottle
– Fill 2L TAE Buffer mixing
bottle to the 1400ml line
with water (tap or distilled)
• Pour 1X TAE into
electrophoresis boxes
stopping about 1cm below
MAX line
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Running gels
• After students have loaded
their gels carefully slide
each gel half back into each
gel tray (in same direction)
and place into the
electrophoresis boxes
– Keep track of which groups’
gels are where!
• Make sure the well sides of
the gels are on the BLACK
electrode side
– Back to Black, RUN to RED
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Running gels
• Top off each box as
needed with TAE to
bring level to MAX
line
• Place lid on each
electrophoresis box
making sure that the
black electrode is at
the well end of the
gels
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Running gels
• Connect the electrodes from
each box to the power supply
and turn on the power by the
switch in the back
• Make sure the power supply
is set on volts and adjust the
voltage to 200*
• When you are ready to start
the run simply press the
button in the center
– Watch for bubbles in the
electrophoresis boxes!
*may need to lower voltage
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After Gel Run
• When the run is complete
(colors have separated) turn off
the power by pressing the
button on the left
• Remove the lids from the
electrophoresis boxes
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After Gel Run
• Carefully remove gel trays
from the box and depending on
time:
– Place each ½ gel on a flat tray on
a patty pac and give back to
groups to analyze
– OR place each ½ gel on a labeled
patty pac and store back in
storage container in refrigerator
until next class meeting and then
distribute on flat trays
• WARNING - WET GELS
ARE VERY SLI PPERY! !
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Next period and so on…
• You can prepare per 2 gels for distribution while
per 1 gels are running and so on…
• Running TAE buffer is good for all classes – no
need to replace unless it gets too hot
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Clean up
• At end of day used buffer can just be flushed
down sink
– Rinse boxes and let air dry
• Used gels can be placed in general trash
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