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15 electrophoresis

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15 electrophoresis 15 electrophoresis Presentation Transcript

  • Gel Electrophoresis gel electrophoresis – moving DNA through a gel medium using an electric current Liquid solutions of the gel is poured into a mould and allowed to set and solidify. Why can we move DNA with electricity? DNA has a negative charge.
  • positive electrode negative electrode agarose gel
  • Gel Electrophoresis Each well is loaded with a different sample of DNA. Samples often contain multiple pieces of DNA of different lengths.
  • Gel Electrophoresis The gel provides resistance for DNA movement. Short DNA moves through gel easily  travels further in a set amount of time  Long DNA requires more effort to move through gel  does not move as far in a set amount of time 
  • Gel Electrophoresis Describe the DNA fragment lengths that have been separated.
  • Gel Electrophoresis The gel medium can be made from: 1. agarose - seaweed extract 2. polyacrylamide - artificial polymer The type of gel used is dependent on how well separated the DNA pieces need to be (resolution). Polyacrylamide has higher resolution than agarose.
  • Gel Electrophoresis DNA is colourless. How do you know that it gets into the gel? Coloured dyes are mixed with DNA to track distance travelled. How do you see where the DNA is after the separation is complete? DNA is stained with ethidium bromide and a UV light box to see fluorescent DNA bands
  • Gel Electrophoresis How do you know how long your DNA fragments are?
  • Gel Electrophoresis A DNA ladder is also loaded on the gel containing known lengths of DNA. Samples of unknown length can then be approximated through comparison with the DNA ladder. The ladder is also known as a DNA marker.