IGEM:MIT/2006/Notebook/2006-9-1

to do

1. glycerol: all LCs -done
2. start a smell test: add precursor to the LCs of 0-Q-119 and O-Q-199 -done, however, results don't look good
3. am miniprep (12): J45397 (1), J45398 (1), J45399 (3), J45299 (3), J45319 (3), pE3 (1) -done
4. analyze: incoming 20 sample sequencing order -done
   • throw out bad sequences -done
5. digest: J45299 (3) and J45319 (3) with XP -done
   • run a gel
6. sequence: (32 wells) -done
   • J45220-C-mut with ATF1 primers -done
   • all other minipreps (except pE3) -done
   • J45398-no mut: 3,4,6,7 -done
7. PCR: pmsCEAB out of pE3(new) and pE3R -done
   • adjust annealing time, check GC content, ask advisor advice on PCRing large coding regions -done
   • clean up??
   • run a gel
8. graph the plate reader data (call stephen)
9. 3-part ligate/transform into A/C backbone:
   • J45299:XP with R0011:ES
   • J45319:XP with R0011:ES
10. pellet: yeast cells in big centrifuge, resuspend in milk, try baking
    • ask samantha for help
11. pm miniprep: iGEM yeast
    • follow yeast miniprep protocol, vortex cells first, ask samantha for help
    • we can do XS digest/freeze of yeast miniprep on monday and ligate/transform with ATF1 when samantha is around and when we have tried out the bread machine