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Team:Caltech/Week 4 - 2010.igem.org
 

Team:Caltech/Week 4

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** Plated 100uL on LB-Tet agar plate & inoculated a 5mL LB-Tet overnight culture.
** Plated 100uL on LB-Tet agar plate & inoculated a 5mL LB-Tet overnight culture.
* Inoculated 5mL LB-Kan culture of purely [http://partsregistry.org/Part:BBa_V1012 V1012] in case we need to retry the competence procedure.
* Inoculated 5mL LB-Kan culture of purely [http://partsregistry.org/Part:BBa_V1012 V1012] in case we need to retry the competence procedure.
 +
* Tested pigment production of cells containing [http://partsregistry.org/Part:BBa_K274100 K274100] in various solutions.
 +
** Tested solutions of LB, LB without yeast extract added, 80% glycerol, 100% glycerol, and glycerol with different concentrations of agar.
 +
** Since the brick is under an arabinose-inducible promoter, two versions of each solution were made - one containing arabinose, and one without arabinose.
 +
** Pigment production was subjectively measured every hour for five hours.
==Thursday 7/8==
==Thursday 7/8==

Revision as of 19:54, 8 July 2010


iGEM 2010



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Contents

Monday 7/5

Note: Today was another Institute Holiday.

Tuesday 7/6

  • Hydrolyzed linseed oil. The reaction solution will be characterized on Wednesday.

Wednesday 7/7

  • Attempted to make V1012 (strain CP919, KanR, envZ deficient) electrocompetent:
    • Centrifuged overnight 3mL LB-Kan culture at 4C, 16000rcf for 10min.
    • Discarded supernatant. Resuspended pellet in 1.5mL ice-cold water. Centrifuged again.
    • Repeated with 0.75mL ice-cold water.
    • Resuspended pellet in 1.5mL ice-cold 10% glycerol.
    • Flash-froze 100uL aliquots in dry ice/ethanol.
  • Transformed light/lysis ligation product from last week into V1012 via electroporation. (Voltage: 2.5V; time constant: 4.5)
    • Plated 100uL on LB-Tet agar plate & inoculated a 5mL LB-Tet overnight culture.
  • Inoculated 5mL LB-Kan culture of purely V1012 in case we need to retry the competence procedure.
  • Tested pigment production of cells containing K274100 in various solutions.
    • Tested solutions of LB, LB without yeast extract added, 80% glycerol, 100% glycerol, and glycerol with different concentrations of agar.
    • Since the brick is under an arabinose-inducible promoter, two versions of each solution were made - one containing arabinose, and one without arabinose.
    • Pigment production was subjectively measured every hour for five hours.

Thursday 7/8

  • The V1012 transformation appears to have worked - the plate is covered in 1000+ colonies.
    • Made freezer stock from parallel liquid culture.
  • Miniprepped the DNA for further modification and for sequencing.


Friday 7/9

Weekend 7/10-11

 
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