User:Tara K. Luckau/Notebook/Team ConGen/2011/06/24

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Contents


SCOC Multiplex 1

PCR

  • multiplex looks good in some PCRs (when run next to uniplexes), but not in others (when scaled up for frag submission)
  • try full gradient (temperature and buffer) of multiplexed reaction


Gel

Pour Load Run
270 mL 1x TAE + 5.4 g agarose 1 µL gel load dye + 5 µL PCR product 160 V
27 µL GelRed 6 µL ladder 50 minutes


  • not favorable results
  • lane for Buffer E, 64.3° consistent with 23 June 2011 PCR/gel
  • in most lanes, can see the Scun9/10/11 band
  • some lanes tend to have larger (Scun15, 22) or small amplicons (Scun2, 3)


  • given that this doesn't seem to be able to get 'better', will submit yesterday's PCR to UAGC
  • should be able to see which loci aren't amplifying, which loci are dominant


  • might be able to split the multiplex into two separate multiplexes
    • looking at Gradient Summary, 13 June 2011:
    • Scun3, 9, 11 - Buffer D, 64°C
    • Scun2, 10, 15, 22 - Buffer D, 56°C
    • Scun10, 15, 22 - Buffer G, 61°C



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