Gel Electrophoresis
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Transcript of Gel Electrophoresis
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Gel ElectrophoresisBy: Brian Barnes, Gary Pope, Eliza Morgan
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What is GE?
• Pronounced Gel ee-LEK-tro-fo-REE-sis• Technique for separating DNA or Protein
Molecules into strands according to length
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What’s The Point?
• Used to separate DNA and identify strands of interest
• Strands of interest are further analyzed once identified
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The Process• Test tube contains DNA• Samples put in different “wells”• Each well contains unique enzyme
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The Process• Gel Matrix, usually agarose• Blue tracking dye added for identification
purposes• Electric current applied to move negatively
charged DNA through agarose
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• Agarose like a spongy barrier• Shorter strands of DNA move more freely
through the gel• Longer strands have more difficulty moving
The Process
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The Process
• Blue Dye is visible moving through gel• As dye touches the end, current is turned off
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The Process• Fluorescent dye is added to identify molecules• Size Standards Well contains known DNA
fragments to compare sizes• Measured by Base Pairs (bp)
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The Process
• Each band is comprised of millions of fragments of the same size
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Analysis
• Now to isolate fragments of interest…• The Blot- a copy of the results which enables
the researcher to identify fragments of interest
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Analysis
• Gel is placed in basic solution• DNA denatures into single strands rather than
double helix
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Analysis• Transferred to salt solution• Nylon Filter placed on top• Absorbent paper towels placed on filter
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Analysis• Salt solution drawn upward by paper towels• DNA adheres to nylon filter• Filter is treated, DNA adheres permanently
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Analysis• Radioactive DNA “probe” added• DNA probe complimentary to band of interest• Probe hybridizes with bands of interest
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Analysis• Filter washed to remove un-hybridized DNA• X-Ray film placed over filter• Radioactive probe exposed under X-Ray
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Using the Analysis• Now the strands of interest are identified• Identical gel is made• DNA of interest is cut out for further analysis