protocol for agar electrophoresis
TRANSCRIPT
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PROTOCOL FOR
AGARELECTROPHORESIS
Jose Manuel Bocara
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MATERIAL REQUIRED FOR THE PROCESS
1 g Agar 10 ml of TAE 50x1 Erlenmeyer of at
least 100 ml1 Bottle/recipient of at least 100 ml
1 Scale (not provide with the kit)
1 Microwave or hot plate
Paper tissues or gloves (not provide
with the kit)
500 ml of destilled water (not provide
with the kit)
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MATERIAL NEEDED FOR EACH PAIR OF
STUDENTS
1 Gel chamber
1 Comb with at least 4 wells
2 Carbon papers aprox. 4x1.5 cm
2 Electric cables: 1 black, 1 red
3x9 V batteries or a power supply for electrophoresis
1 Syringe 4x10 ul tips
4 Tubes with coloured samples
1 Piece of black and white paper (A5 laminated)
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PROCESSES OF ELABORATION
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Prepare the TAE 1x solution
Dilute 10 ml of TAE
(concentration 50x) in 490 ml of
destilled water
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Prepare your electrophoresis box
• Put the comb with 6
wells in the slot.
• Place your box on the
black paper
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Prepare the Agar gel at 1%
• Weigh out 1 g of agar powder.
• Mix it with 100 ml of TAE 1x.
• Heat it in the microwave or on
a hotplate, until the solution
becomes transparent
1 2
3
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A DIAGRAM
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Load the samples
1. Cut off the gel that has gone over the litle wells using a ruler.
2. Put carbon paper over each end of the box.
3. Pour TAE 1x solution over the gel to cover it.
4. Remove the comb slowly and in a vertical position.
5. Get the syringe and put a 10 ul tip on the end.
6. Take out spproximately 10 ul of the first solution (to the second line marked on the tip).
7. Put the samples in the first well (or as was decided in point 4). Ensure your hand doesn't
shake.
8. Load the other samples into the other wells and do not forget to change your tip each time
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Load of Samples images
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Run the Gel
Connect 3.9 V batteries in series
Or set the power supply at 50 V
Connect the cables, using the colour code:
Black-negative side on the top, close to the samples Red-positive
side, at the bottom, far away from the samples.
Turn on the power (run button)
Wait approximately 15 minutes, until different
bands appear.
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Run the Gel images
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Observation
The samples were composed
of different colours, each
colour moves at a different
specific speed in the gel,
attracted by the positive
pole.
S1 S2S3EC
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Result
Place your box on white
paper so you can see the
result more clearly
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Conclusion
• Electrophoresis is a technique that allows us to separate
components by size
• We can see that some of the samples have 2 components and others
have 3.
• The yellow component is the smallest and is the most negatively
charged as it moves the furthest through the gel towards the positive
anode.
• The blue components is the largest and has the least negative charge
as it moves the least through the gel.
Conclusion:
Sample S3 has the same
components as the crime
Scene (EC) sample.
S1 EC S3 S2
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END OF PROTOCOL
Jose Manuel Bocara