Friday 11/09/12

9:30 A – 8:30P

  • Making double mutants
  • Collect virgin TM2/TM6 flies
  • Clear Pr/TM3,hbZ and Sp/CyOZ for virgin collection to remake double balanced 2,3 flies.
  • TM6b = AntpHue1 .
  • TM2 = Ubx130esemc2 .
  • E(spl) and sim[D](7646) both have TM6b, Tb.  Should be lethal with TM6.    First cross of E(spl) x sim[D] screening against Hu and Tb should have been sufficient to get only true recombinants.
  • Next cross E(spl)/sim[D]   x   TM2,Ubx/TM6,Tb.  The humoral guys should have TM6 and be tubby still. The non-humoral guys should have a TM2 balancer.   Most of these will be E(spl)/TM or sim/TM.  some fraction will be +/TM and some fraction should be Espl,sim/TM.    These will all look the same.   Cross them to TM2,Ubx/TM6,Tb.  Double balanced guys in the next generation will be ebony.  Non-ebony flies will be a all the same ?-allele / TM2 or TM6.  These can now be safely selfed without mixing different ?-alleles, until there are enough ?-alleles to grind up embryos.  Then we need to be able to PCR across the gaps –> need mapped gaps.
  • Determining primers for E(spl) locus testing:  Lpr1 upstream (~96F2 = chr3R:21600K to 97A=21900K (300Kb region).  BSC751 is a smaller deletion with mapped ends, would have been a better choice. Still kills entire Espl complex + Gro gene.    Maybe should try this?
  • PM virgin collection

Meeting with Welcome Bender (11:00A – 1:30P)

OligoPaints website (from Brian)

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