IGEM:MIT/2006/Notebook/2006-9-8

to do

1. start LCs for plate reader?? (SP ?)
2. check sequences (dye blob re-runs should be in from 9/1 order)
3. glycerol any of the LCs in fridge that look good post this second sequence analysis
   • discard of confirmed bad dna and glycerols!
4. glycerol 6 new LCs (3 of the 399-1s and 3 of the 320-Cs)
5. Y0080 and Y0078 yeast vectors
   • miniprep
   • digest with XS --->must do the artic freeze step following digest
     • run results on a gel
   • ligate to J45014:XS (already cut and ready to go in corner of digest tray)
   • transform
6. pmsCEAB
   • digest with ES (has 4 Pst sites!! i said something about Eco in an email, but they are definately Pst!)
     • run results on a gel and clean-up
   • ligate into a double resistance backbone:ES (make sure different from w-ever our WGD device is living in)
   • transform
7. pellet new yeast cells (pm) and resuspend in milk -- bake again or bake on saturday (KB in pm)

Transformation of J45400

• ligated J45399 (4 of them...1A, 1B, 1C, and 2A) to R0011, in A/T backbone, to make J45400 in A/T backbone. We still need to check the sequencing of the J45399s to see which ones were right...hopefully one of the 4 I used was correct. there wasn't a lot of R0011, so I chose 1A, 1B, and 1C b/c they looked the best on the gel, and 2A for the heck of it.

Osmy+E0840 for plate reader

• transformed osmy+E0840 FN for the plate reader...used an amp plate b/c I wasn't sure what the resistance was.