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Gel Electrophoresis

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The 'gel' in gel electrophoresis is the component that physically separates ... move through the gel toward the positive pole (Remember: Opposites do Attract! ... – PowerPoint PPT presentation

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Title: Gel Electrophoresis


1
Gel Electrophoresis
Developed by Science Behind Our Food University
of Georgia Adapted by Georgia Ag Ed Curriculum
Office, 2004
2
What is Gel Electrophoresis?
  • Electrophoresis Movement of charged particles
    through a solution (gel) under the influence of
    an electric field
  • DNA fragments have a negative (-) charge and will
    move toward the positive electrode

3
The Basic Components
  • The gel in gel electrophoresis is the component
    that physically separates the molecules.
  • The electric power supply provides the
    electricity that carries the molecules through
    the gel
  • The electrophoresis chamber is where it all takes
    place.

4
The Process
  • Gels are often made of a gelatin-like substance
    called agarose.
  • A comb with teeth is suspended in the gel until
    it cools.

5
  • When the gel has cooled and solidified, the
    scientist pulls the comb out of the gel.
  • The teeth of the comb leave impressions in the
    gel called wells where the DNA will be loaded.

6
  • The gel is then placed in the electrophoresis
    chamber.
  • Gel acts like a bridge through which the
    molecules must travel to get from one side of the
    chamber to the next.

7
DNA PreparationRestriction Enzymes
  • Cut DNA into smaller pieces
  • These special enzymes recognize specific
    sequences in the DNA molecule (for example
    GATATC) and cuts the DNA wherever that sequence
    occurs.

8
Loading DNA Samples into the Gel
  • Gels usually contain several different samples of
    DNA for study.
  • The researcher uses a pipette to draw out a
    small, precisely measured amount of each sample
    and places each sample in its own well.

9
The electrophoresis After all the DNA samples
have been loaded into the wells, the electrodes
are connected to the power supply. When the
power is turned on, the negatively charged
molecules move through the gel toward the
positive pole (Remember Opposites do Attract!!).
Both the DNA fragments and the dye molecules have
a negative charge.
10
http//www.life.uiuc.edu/molbio/geldigest/electro.
htmlrun
11
http//www.life.uiuc.edu/molbio/geldigest/electro.
htmlrun
12
(No Transcript)
13
Applications of Gel Electrophoresis
  • DNA
  • Criminal investigations
  • Blood
  • Saliva
  • Hair
  • Semen
  • Paternity Tests

14
Forbidden Donut Mystery
  • Your Analysis Unit is charged to determine who
    took a bite of Homers Forbidden Donut.
  • Posterboard ?
  • Names on the Posterboard ?
  • Scissors ?
  • DNA strips from suspects

15
Forbidden Donut Mystery
  • Your Analysis Unit is charged to determine who
    took a bite of Homers Forbidden Donut.
  • Posterboard GEL
  • Names on the Posterboard ?
  • Scissors ?
  • DNA strips from suspects

16
Forbidden Donut Mystery
  • Your Analysis Unit is charged to determine who
    took a bite of Homers Forbidden Donut.
  • Posterboard GEL
  • Names on the Posterboard WELLS
  • Scissors ?
  • DNA strips from suspects

17
Forbidden Donut Mystery
  • Your Analysis Unit is charged to determine who
    took a bite of Homers Forbidden Donut.
  • Posterboard GEL
  • Names on the Posterboard WELLS
  • Scissors RESTRICTION ENZYME
  • DNA strips from suspects

18
Forbidden Donut Mystery
  • How to cut and place sequences on gel

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