15 electrophoresis-2010 update

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15 electrophoresis-2010 update

  1. 2. Gel Electrophoresis <ul><li>gel electrophoresis – moving DNA through a gel medium using an electric current </li></ul><ul><li>Why can we move DNA with electricity? </li></ul><ul><li>DNA has a negative charge. </li></ul>
  2. 3. Vertical Gel http://www.siumed.edu/~bbartholomew/images/chapter6/F06-21.jpg
  3. 4. negative electrode positive electrode agarose gel
  4. 5. Gel Medium <ul><li>The gel medium can be made from: </li></ul><ul><li>1. agarose - seaweed extract </li></ul><ul><li>2. polyacrylamide - artificial polymer </li></ul><ul><li>Liquid solutions of either medium are poured into a mould and allowed to set and solidify. </li></ul>
  5. 6. Gel Medium <ul><li>The semi-solid provides resistance for DNA movement. </li></ul><ul><li>Short DNA </li></ul><ul><ul><ul><li>moves through agarose easily </li></ul></ul></ul><ul><ul><ul><li>travels further </li></ul></ul></ul><ul><li>Long DNA </li></ul><ul><ul><ul><li>requires more effort to move through agarose </li></ul></ul></ul><ul><ul><ul><li>does not move as far </li></ul></ul></ul>
  6. 7. http://cg.scs.carleton.ca/~morin/teaching/compbio/f_s02gelelect.gif
  7. 8. DNA Visualization <ul><li>DNA cannot be observed in gel </li></ul><ul><li>coloured dyes mixed with DNA to track distance travelled </li></ul><ul><li>stain DNA with ethidium bromide </li></ul><ul><li>UV light box to see fluorescent DNA bands </li></ul>
  8. 12. Gel Electrophoresis <ul><li>http://www.sumanasinc.com/webcontent/animations/content/gelelectrophoresis.html </li></ul>

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