Agarose Gel Electrophoresis

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Agarose Gel Electrophoresis

• 2002. 1.22.
• Park, Woosin

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1. Principle

• DNA is cut up with restriction enzyme
• The DNA fragments are transferred
• through wells cut into a gel
• An electric charge is transferred through
• the gel. The negatively charged DNA
• fragments are repelled by the negative
• electrode and attracted to the positive
• electrode.
• As the size of fragment increases, the band
• appears near the negative electrode

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2. Agarose gel

• Agarose gel conc. According to the size of DNA

Selected in this experiment
2.0 2g agar 100 ml TAE buffer
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3. Making a gel plate (2.0 )

• Dissolve 2.0 g of agar powder into 100
• ml of TAE buffer
• Heat the mixture in a electric range at 600 w
• for 2 min
• Spread the solution 25 ml on a big plate and
• 10 ml on a small plate respectively and leave
  
• them in the air for 15 min
• Wells in the gel would be formed by using a comb

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• Solidified agarose gel

Max. sample vol. 10 µl
Max. sample vol. 8 µl
Small gel plate
Big gel plate
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4. Reagents

• DNA Ladder
• standard DNA showing the position of
• bands according to their lengths after
• electrophoresis
• Loading solution deep blue color
• Loading solution 1.8 µl (2.0 for big wells)
• sample 8 µl (10 for big wells) and 8 µl (10
• for big wells) of the mixture into each well
• One blank sample should be involved

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Length
long
Size determination
short
Result of electrophoresis of DNA ladder
Sample 1
Sample 2
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5. Condition for electrophoresis

• 100 v for 35 min or 50 v for 70 min

6. Dying

• Solution 2 µl Ethidium bromide TAE
• 200 ml
• Submerge the gel in the solution and
• shake very slowly for 20 -30 min

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6. Trans-illumination with U.V.7. Take a picture