User:Stuart McKellar/Notebook/Bird Sex Testing/2012/10/19

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Electrophoresis Test

Objective: Try and visualise DNA on an agar gel. Method: Poured a 2% Gel pre-stained with GelRed 1:10,000. This was from a while ago so I may need to post stain it. Heated in microwave for a couple of minutes and then let cool. Poured gel, added 3 x 3.3ul of 300-10KB ladder. Added 1 x 5ul ladder in 4th lane. Ran gel at 60ma and 100V (volts as recommended).

Results: DNA had visibly moved in first few minutes. Gel box underneath was on but no visualisation yet. I may be using the wrong dye for this gel box but Gel Red should have an excitation peak at the wavelength. I am not sure what wavelength it emits at though.

No visible results. This could be for the following reasons:

  • The ladder is degraded
  • The GelRed is not emitting light at a visible frequency
  • There is not enough contrast
  • The buffer is at an incorrect pH and the DNA is not moving with the bromophenol blue

And that is all the reasons that I can think of.



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