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

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(to do)
(to do)
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==to do==  
==to do==  
-
#glycerol: all LCs
+
#glycerol: all LCs '''-done'''
-
#start a smell test: add precursor to the LCs of 0-Q-119 and O-Q-199
+
#start a smell test: add precursor to the LCs of 0-Q-119 and O-Q-199 '''-done, results don't bode well though'''
-
#am miniprep (12): J45397 (1), J45398 (1), J45399 (3), J45299 (3), J45319 (3), pE3 (1)
+
#am miniprep (12): J45397 (1), J45398 (1), J45399 (3), J45299 (3), J45319 (3), pE3 (1) '''-done'''
#*note: I don't think that the yeast will be grown up enough to miniprep this early in the day
#*note: I don't think that the yeast will be grown up enough to miniprep this early in the day
-
#analyze: incoming 20 sample sequencing order
+
#analyze: incoming 20 sample sequencing order '''-done'''
-
#*throw out bad sequences
+
#*throw out bad sequences '''-done'''
-
#digest: J45299 (3) and J45319 (3) with '''XP'''
+
#digest: J45299 (3) and J45319 (3) with '''XP''' '''-done'''
#*run a gel
#*run a gel
-
#sequence: (32 wells)
+
#sequence: (32 wells) '''-done'''
-
#*J45220-C-mut with ATF1 primers
+
#*J45220-C-mut with ATF1 primers '''-done'''
-
#*all other minipreps (except pE3)
+
#*all other minipreps (except pE3) '''-done'''
-
#*J45398-no mut: 3,4,6,7
+
#*J45398-no mut: 3,4,6,7 '''-done'''
-
#PCR: pmsCEAB out of pE3(new) and pE3R  
+
#PCR: pmsCEAB out of pE3(new) and pE3R '''-done'''
-
#*adjust annealing time, check GC content, ask advisor advice on PCRing large coding regions
+
#*adjust annealing time, check GC content, ask advisor advice on PCRing large coding regions '''-done'''
#*clean up??
#*clean up??
#*run a gel
#*run a gel
Line 20: Line 20:
#*J45299:XP with R0011:ES
#*J45299:XP with R0011:ES
#*J45319:XP with R0011:ES
#*J45319:XP with R0011:ES
-
#*need to find more backbone!!
 
#pellet: yeast cells in big centrifuge, resuspend in milk, try baking  
#pellet: yeast cells in big centrifuge, resuspend in milk, try baking  
#*ask samantha for help
#*ask samantha for help

Revision as of 15:00, 1 September 2006

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, results don't bode well though
  3. am miniprep (12): J45397 (1), J45398 (1), J45399 (3), J45299 (3), J45319 (3), pE3 (1) -done
    • note: I don't think that the yeast will be grown up enough to miniprep this early in the day
  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
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