User:Paul Rothenberg/Notebook/Pauls Notebook/2013/02/25

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25 February 2013

  • Objectives:
  • 1. Create a 1% agarose gel (1g agarose in 100 mL TAE) for gel electrophoresis
  • 2. Run a DNA sample to test for expression

  • Procedure:
  • 1. 50x TAE stock buffer was diluted to 1x TAE buffer, using 2 mL of 50x buffer and 98 mL of ultra-pure water (1)
  • 2. 0.25 grams of agarose was dissolved in 25 mL of TAE
  • 3. The TAE was microwaved for 45 seconds to dissolve the agarose
  • 4. The TAE and agarose was poured into the electrophoresis chamber and left to harden; electrophoresis buffer was added following
  • 5. 5 microliters of DNA and 1 microliter of dye; 6 microliters of the DNA ladder were added to the wells
  • 6. The gel was run for 55 minutes at 90V
  • 7. The gel was then placed in Ethidium Bromide rinse and shaken for 15 minutes
  • 8. The gel was removed and placed in a TAE rinse and shaken for another 15 minutes
  • 9. The gel was exposed in an electrophoresis chamber

  • Data:
  • 1. No experimental DNA bands were visible under UV radiation. The gel will either have to be stained in ethidium bromide for a longer period of time, or a new gel will have to be rerun.