Team:Caltech/Week 4

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Revision as of 00:57, 8 July 2010

iGEM 2010

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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, <math>\Delta</math>envZ) 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.

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@@ Line 10: / Line 10: @@
 ==Wednesday 7/7==
+* Attempted to make [http://partsregistry.org/Part:BBa_V1012 V1012] (strain CP919, KanR, <math>\Delta</math>''envZ'') 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.
 ==Thursday 7/8==