User:Tara K. Luckau/Notebook/Team ConGen/2011/10/31

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SCOC MP1

  • First time running multiplex within lanes
  • First time post-PCR mixing (1:1:1)
  • First time sub-sampling across San Diego county, focusing on RAJ/HOL and SYR
  • will send individual MP plates (blue, green, yellow) to Tucson, in case mixing doesn't work out
  • UAGC may be able to play with ratios
  • UAGC will be able to run one color at a time, if needed


PCR

  • 20111031 PCRa.png
  • oops - mispipetted (dispensed, but aspirated before pulling tip out of PCR plate) SCOC RAJ 49, redispensed into SCOC RAJ 48
SCOC RAJ 48 may be contaminated with SCOC RAJ 49
SCOC RAJ 49 may have low amount of template DNA
  • had lots of left-over master mix ... decrease to 5% overage (from 10% overage) to save on reagent cost


  • 20111031 PCRb.png
  • still had lots of left-over master mix! could decrease even more, to 3% overage


  • 20111031 PCRc.png
  • still had lots of left-over master mix! could decrease even more, to 3% overage


1Kb Ladder

  • made two aliquots of 1Kb ladder, according to protocol
120µL ladder + 880µL H2O + 200µL loading dye = 1200µL ladder mix


dNTP MIX

  • made dNTPs according to protocol
125µL dATP + 125µL dTTP + 125µL dCTP + 125µL dGTP + 4500µL H2O = 5000µL dNTP Mix @ 10mM (2.5mM each dNTP)
makes 5 aliquots