User:Tara K. Luckau/Notebook/Team ConGen/2011/06/13

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Contents


Gradient Summary

  • favorable conditions (temperature and buffer) is indicated by green highlighting, and corresponds to the green happy faces on each gel
  • right-most column indicates number of primer pairs that work well under that condition
  • Image:20110613 GradientSummary.jpg
  • will next try to multiplex 7 primer pairs using Buffer E at 64°C
    • Scun2
    • Scun3
    • Scun9
    • Scun10
    • Scun11
    • Scun15
    • Scun22


Multiplex 1

  • The first attempt at multiplexing!

PCR

  • reaction conditions are based on the gradient summary (above) - Buffer E, 64°C
  • run uniplexes at this reaction condition to check against the gradient PCR result, 4 replicates
  • run multiplex at this reaction condition, 8 replicates
  • hopefully, we'll get clear bands in both uniplexes and multiplex!

Gel

  • Expected banding (based on gradient PCRs)


  • Conclusion: missing the Scun22 band, but the other 6 look like they might be there
  • ready to order labelled primers, and start multiplexing and fragging!



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