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

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(to do)
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#*transform
#*transform
#pellet new yeast cells (pm) and resuspend in milk -- bake again or bake on saturday (KB in pm)
#pellet new yeast cells (pm) and resuspend in milk -- bake again or bake on saturday (KB in pm)
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==Transformation of J45400==
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*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.
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==Osmy+E0840 for plate reader==
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*transformed osmy+E0840 FN for the plate reader...used an amp plate b/c I wasn't sure what the resistance was.

Revision as of 16:32, 8 September 2006

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.
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