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)