IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/06/21: Difference between revisions

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[[Image: Ltp bet ger 621.jpg|thumb|left| lanes 2,11,12 were successful]]
[[Image: Ltp bet ger 621.jpg|thumb|left| lanes 2,11,12 were successful]]


==Team Genetic Fence==
==Team Genetic Fence==

Revision as of 09:12, 22 June 2010

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Team Allergy

PCR (see gel result below)

lanes 2,11,12 were successful






Team Genetic Fence

Barnase and Barstar

Barnase and Barstar Plasmids from ADDGENE arrived in mail

pMT316 - Barstar. Info from ADDGENE

pMT413 - Barnase, Barstar. Info from ADDGENE

pMT1002 - Barnase, Barstar. Info from ADDGENE


Notes from lab meeting

LVA tail for Cre


GAL4 DBD Innoculation

Pipetted 5mL of LB+Amp each into capped cc tubes, and mixed bacterial scrapings from 5 separate colonies of transformed GAL4 DNA binding domain E.Coli. Set to shake at 37°C overnight.


Cre, Lox66, Lox 71, GFP, and Luciferase Bacterial Transformation

Performed bacterial transformation according to Silver:_Bacterial_Transformation, however we used 1μL in step 3, and used entire tube of TOP10 competent cells instead of only 10-15μL.


Transformed the following each into its own tube of TOP10 E.Coli:

Cre DNA Recombinase

  • biobrick BBa_J61047, located on plate 1, well 5D, plasmid pSB1A2

Lox71

  • biobrick BBa_I718017, located on plate 1, well 17J, plasmid pSB1A2

Firefly Luciferase

  • biobrick BBa_I712019, located on plate 1, well 10H, plasmid pSB1AK8.

GFP mutant (phenotype identical to wildtype)

  • biobrick BBa_E0040, located on plate 1, well 14K, plasmid pSB1A2.

Lox66

  • biobrick BBa_I718016, located on plate 1, well 17H, plasmid pSB1A2.


Labeled each plate LB+Amp, its contents (ie Lox71 or Firefly Luciferase), MP, JW, OM, 6-21-10

Set all five plates to incubate at 37°C overnight.




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