Team:Caltech/Week 4

(Difference between revisions)

Revision as of 01:33, 9 July 2010

iGEM 2010

People

Project Details

Protocols

Completed Systems

Notebook

Biosafety

Human Impact

References

Support

Note: Today was another Institute Holiday.

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

Attempted to make [http://partsregistry.org/Part:BBa_V1012 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 [http://partsregistry.org/Part:BBa_V1012 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 [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.

The [http://partsregistry.org/Part:BBa_V1012 V1012] transformation appears to have worked - the plate is covered in 1000+ colonies.
- Made freezer stock from parallel liquid culture.
Miniprepped the DNA and prepared a sample for sequencing.
Started LB liquid culture of [http://partsregistry.org/Part:BBa_M30109 M30109] for assembly tomorrow, along with cultures of [http://partsregistry.org/Part:BBa_J04450 J04450] to perform a quick lysis test (in preparation for testing the lysis construct).
Prepared 2YT and SOB media in preparation for making electrocompetent DH5\alpha cells tomorrow. Also prepared all glassware and other supplies.
Completed schematics for our printing process, including a detailed gene network diagram. We will begin assembly of the 3D genetic constructs tomorrow.

Error creating thumbnail: /websites/2010.igem.org/wiki/bin/ulimit4.sh: line 4: convert: command not found

@@ Line 25: / Line 25: @@
 ==Thursday 7/8==
-* The V1012 transformation appears to have worked - the plate is covered in 1000+ colonies.
+* The [http://partsregistry.org/Part:BBa_V1012 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.
+* Miniprepped the DNA and prepared a sample for sequencing.
+* Started LB liquid culture of [http://partsregistry.org/Part:BBa_M30109 M30109] for assembly tomorrow, along with cultures of [http://partsregistry.org/Part:BBa_J04450 J04450] to perform a quick lysis test (in preparation for testing the lysis construct).
+* Prepared 2YT and SOB media in preparation for making electrocompetent DH5\alpha cells tomorrow. Also prepared all glassware and other supplies.
+* Completed schematics for our printing process, including a detailed gene network diagram. We will begin assembly of the 3D genetic constructs tomorrow.
 ==Friday 7/9==