Difference between revisions of "User:Stuart McKellar/Notebook/Bird Sex Testing/2013/01/15"

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neg
 
neg
  
 
+
Results: no bands. Try lower annealing temperature - say 55°C
  
 
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Revision as of 00:16, 18 January 2013

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Results

  • last samples produced valid results when compared with positive controls

Repeating experiment with 2550/2870 primer pair. Compare the photos and decide 100% which set I will be using.

Method: run a PCR @ 60°C and with 1ul 2550/2870 primers. Visualise on a 1% gel. 10 samples.

Master Mix: 7 x lorikeets 2 x positive controls (male female) 1 negative control

  • 1ul F primer
  • 1ul R primer
  • 2.0ul dNTPs
  • 0.5u Taq
  • 1ul DNA
  • 2.5ul 10x buffer
  • 1.5ul MgCl2
  • 16ul H2O
  • 1:1SWWBD
  • 2:2SWWBD
  • 3:R5#9
  • 4:R5#51
  • 5:R5#52
  • 6:R5#100
  • 7 :Pink Gold

male female neg

Results: no bands. Try lower annealing temperature - say 55°C