User:Stuart McKellar/Notebook/Bird Sex Testing/2012/11/13

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gel electrophoresis

  • The aim of this lab is to determine the sex of the rainbow lorikeets through agar gel electrophoresis. Gel is 2% agar and stained with GelGreen that has been sitting on my desk room temperature since last test (which was successful). I will run a ladder as well to make sure that the gel is still showing luminescence.

If the PCR failed, I expect the DNA not to migrate. I am loading 5ul DNA sample with 1ul x 6x DNA running buffer (glycerol + bromophenol blue).

Samples were loaded as follows:

  • 1SWWBD
  • 2SWWBD
  • R5#9
  • Ladder (300-10KB)
  • R5#51
  • R5#52
  • R5#100
  • Pink Gold

Gel ran at 100V (intially started at 300V for a few seconds and then pansied out) and 60mA. Immediate illumination could be seen in the well with the ladder. Gel is running and I can see the ladder but nothing else. Fuck. Looks like I am going back to PCR. After 300bp had reached centre, still no light from samples. Maybe run a single lorikeet sample through PCR and then check on a gel.