User:Tara K. Luckau/Notebook/Team ConGen/2011/06/23

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SCOC Multiplex 1

PCR

  • similar to 13 June 2011 PCR - uniplexes of each primer pair, then multiplex, using labeled primers
  • 20110623 PCR.jpg


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


  • see 13 June 2011 for banding pattern expectations
  • 20110623 Gel.jpg
  • consistent with 13 June 2011 results (missing Scun22 band, but all others seem to be there)
  • notice bright primer band on multiplex only - probably could do some optimization to make reaction more efficient


SCOC Multiplex 1

  • SCOC Multiplex 1 diversity panel with labelled primers to send to frag, part deux!


PCR

  • 20110623 PCR2.jpg
  • some of the samples used for the diversity panel are getting low ... should switch out soon
  • SCOC TP3 X01 was low, like the others, but I already had a SCOC TP3 extracted last year in the Mass Extract ... so used it, SCOC TP3 X06


Gel

  • used left-over gel from 13 June 2011
Load Run
2 µL gel load dye + 4 µL PCR product 160 V
6 µL ladder 1 hour


  • see 13 June 2011 for banding pattern expectations
  • 20110623 Gel2.jpg
  • still a whole lotta nada  :(
  • strong primer band, again
  • a bit of amplification of Scun9/10/11, but it's crap


Next Steps

  • try full gradient on multiplex, ugh