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Gel Electrophoresis

Gel Electrophoresis. By: Brian Barnes, Gary Pope, Eliza Morgan. What is GE?. Pronounced Gel ee-LEK-tro-fo-REE-sis Technique for separating DNA or Protein Molecules into strands according to length. What’s The Point?. Used to separate DNA and identify strands of interest

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Gel Electrophoresis

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  1. Gel Electrophoresis By: Brian Barnes, Gary Pope, Eliza Morgan

  2. What is GE? • Pronounced Gel ee-LEK-tro-fo-REE-sis • Technique for separating DNA or Protein Molecules into strands according to length

  3. What’s The Point? • Used to separate DNA and identify strands of interest • Strands of interest are further analyzed once identified

  4. The Process • Test tube contains DNA • Samples put in different “wells” • Each well contains unique enzyme

  5. The Process • Gel Matrix, usually agarose • Blue tracking dye added for identification purposes • Electric current applied to move negatively charged DNA through agarose

  6. The Process • Agarose like a spongy barrier • Shorter strands of DNA move more freely through the gel • Longer strands have more difficulty moving

  7. The Process • Blue Dye is visible moving through gel • As dye touches the end, current is turned off

  8. The Process • Fluorescent dye is added to identify molecules • Size Standards Well contains known DNA fragments to compare sizes • Measured by Base Pairs (bp)

  9. The Process • Each band is comprised of millions of fragments of the same size

  10. Analysis • Now to isolate fragments of interest… • The Blot- a copy of the results which enables the researcher to identify fragments of interest

  11. Analysis • Gel is placed in basic solution • DNA denatures into single strands rather than double helix

  12. Analysis • Transferred to salt solution • Nylon Filter placed on top • Absorbent paper towels placed on filter

  13. Analysis • Salt solution drawn upward by paper towels • DNA adheres to nylon filter • Filter is treated, DNA adheres permanently

  14. Analysis • Radioactive DNA “probe” added • DNA probe complimentary to band of interest • Probe hybridizes with bands of interest

  15. Analysis • Filter washed to remove un-hybridized DNA • X-Ray film placed over filter • Radioactive probe exposed under X-Ray

  16. Using the Analysis • Now the strands of interest are identified • Identical gel is made • DNA of interest is cut out for further analysis

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