User:Stuart McKellar/Notebook/Bird Sex Testing/2013/01/19: Difference between revisions

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Latest revision as of 22:22, 26 September 2017

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Preliminary results generation

  • I am getting results that Iam confident are usable. This is by comparing them to the positive control samples and getting ratios that are correct.

I will be doing a PCR with P2/P8 primers and then comparing against positive controls with ALL SAMPLES (except loris). Then I will repeat this with the 2550F primer pair at 55°C and comparing all these results. If I can't get consistency, I will order the NP MP primer set.

Method:

black pen - P2 - 60C blue - 2550F - 55C

29 samples


30 samples master mix

Tube

  • 1ul F primer
  • 1ul R primer
  • 2.0ul dNTPs
  • 0.5u Taq
  • 1ul DNA
  • 2.5ul 10x buffer
  • 1.5ul MgCl2
  • 16ul H2O


Master mix

  • 30ul F primer yes
  • 30ul R primer yes
  • 60ul dNTPs yes
  • 3ul Taq (0.5u per tube)
  • 75ul 10x buffer
  • 45ul MgCl2
  • 480ul H2O

added 35ul oil

3min initial melt\

2550 sample

Master mix

  • 30ul F primer yes
  • 30ul R primer yes
  • 60ul dNTPs yes
  • 3ul Taq (0.5u per tube)
  • 75ul 10x buffer yes
  • 45ul MgCl2 yes
  • 480ul H2O yes


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