Difference between revisions of "User:Stuart McKellar/Notebook/Bird Sex Testing/2012/10/19"

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(Autocreate 2012/10/19 Entry for User:Stuart_McKellar/Notebook/Bird_Sex_Testing)
 
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==Entry title==
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==Electrophoresis Test==
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Objective: Try and visualise DNA on an agar gel.
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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).
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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.
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No visible results. This could be for the following reasons:
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* The ladder is degraded
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* The GelRed is not emitting light at a visible frequency
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* There is not enough contrast
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* The buffer is at an incorrect pH and the DNA is not moving with the bromophenol blue
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And that is all the reasons that I can think of.
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Revision as of 05:10, 19 October 2012

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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.