Transcript Gel electrophoresis teacher Edvotek set up

Gel Electrophoresis
Teacher
Instructions
Edvotek
Set Up
12 groups
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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 SB Buffer
for making gels
• Measure 50ml of 20X
SB Buffer stock
solution into the 50ml
conical SB Buffer
measuring tube
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Prepare 1X SB Buffer
for making gels
• Pour the 50ml of 20X
SB Buffer stock
solution into the 1L
SB Buffer mixing
bottle
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Prepare 1X SB Buffer
for making gels
• Fill 1L SB Buffer mixing
bottle to the 1000ml line
with water (tap or
distilled)
• You’ll need to repeat this
as necessary for your
number of classes - each
bottle should prepare
enough 1X SB Buffer for
2 1/2 classes
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Prepare 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 2 bottles
per class
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Prepare gels
• Fill each bottle to the
200ml mark with your
prepared 1X SB
Buffer solution
– Bottles have been prechecked for calibration
• Cap tightly and shake
to mix
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Prepare gels
• Loosen cap 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 full boil
starts
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Prepare gels
• While agar is in
microwave assemble
the gel trays
– Can assemble 24 trays at
a time
• Place rubber gaskets on
each side of the gel
casting tray
• Place comb in end slots
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Prepare gels
• Carefully remove the
HOT bottles from the
microwave and swirl be careful of steam
escaping from the
loose caps!
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Prepare gels
• Check that agar has
fully dissolved or, if
re-melting solidified
agar, that it has all
melted back into
solution
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Prepare gels
• Carefully pour hot
agar solution into the
assembled gel casting
trays (okay to pour
while really hot)
• Fill each tray to top of
rubber gaskets
• Each bottle should fill
6 trays
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How to store prepared gels
• After gels have
solidified, remove the
comb and both gaskets
• Carefully slide gel out
of the tray onto a
“patty pac” paper
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How to store prepared gels
• Each paper will hold 2
gels
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How to store prepared gels
• Place 2 papers with 2
gels 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 rest of gels in
storage container and
place container in
fridge for up to a week
• Each container will
hold 12 gels - 1
container per class!
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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
• Then gently slide each gel
back into a casting tray
– Be sure to slide gels into trays in
same orientation they were cast (wells at notch end of tray)
• Try to keep gels and trays low
and level to prevent accidental
tearing of the gel
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Running gels
• Prepare 1X SB Buffer
solution as before
• You’ll need 2 L of
solution total for both
electrophoresis boxes
• Place tray supports in each
electrophoresis box and
pour in 1L of prepared 1X
SB Buffer into each box
• You only need to do this
before the first class
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Running gels
• After students have loaded
their gels carefully place
each gel tray into the
electrophoresis boxes
• The trays will only fit in
one direction ;)
• Remember to use care that
the gels don’t slide out of
the trays as they are
carried to the boxes!
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Running gels
• Place lid on each
electrophoresis box
making sure that the
black electrode is at
the well end of the
gels
– The electrode wires
inside the box are color
coded as well
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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 70
• When you are ready to start
the run simply press the
button on the far right
– Watch for bubbles in the
electrophoresis boxes!
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After Gel Run
• After the colors have separated
turn off the power supply and
remove the gel box lid
• Carefully remove gel trays
from the box and depending on
time:
– Give back to groups to analyze
– OR place each gel on a labeled
patty pac and store back in
storage container in refrigerator
until next class meeting and then
distribute on individual patty pacs
• WARNING - WET GELS
ARE VERY SLI PPERY! !
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Next period and so on…
• Running SB buffer is good for all classes – no
need to replace unless it gets too hot
• You can put gels into trays for periods 1 and 2,
then after removing completed gels from per 1
trays put gels for per 3 into trays while per 2 is
running and so on…
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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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