User:Tara K. Luckau/Notebook/Team ConGen/2012/11/27

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CNHY MP1a/b Optimization

  • optimize for marker peak height - vary each primer (based on results from 23 July 2012)
  • CNHY MP1a: increase Ai5013, Ai5043, Ai5071
  • CNHY MP1a: decrease Acos5
  • CNHY MP1b: increase Ai5057, Ai5062


Frag Results

  • UAGC submission# YFPQ


MP1a

23 July 2012 this time
Ai5043 did not amplify did not amplify; remove from multiplex
Acos3 amplified well; no changes needed still fine, though seems to decrease with increasing Acos5 & Ai5071
Acos5 amplified with high intensity, pull down a bit 0.04 = too low; 0.06 = inconclusive (failures); 0.08 = too high
Ai5013 amplified with very low intensity; increase concentration second peak never got above 250, regardless of concentration; but in Acos5 assay, noticed second peak reached 400 intensity as Acos5 decreased
Ai5071 amplified with lower intensity; increase a bit best at 0.24, 0.28, 0.32 (200-300 intensity only)


MP1b

23 July 2012 this time
Ai5033 amplified well; no changes needed still fine (no amp for any markers in Ai5057 assay)
Ai5057 did not amplify did not amplify; remove from multiplex
Ai5037 amplified well; no changes needed still fine (no amp for any markers in Ai5057 assay)
Ai5035 amplified well; no changes needed a bit low in Ai5062 assay (no amp for any markers in Ai5057 assay)
Ai5062 amplified but with very low intensity; increase concentration best at 0.48 (400 & 6000 intensity)