Lab Notebook

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'''May 15''': Transformed Top10F' bacteria with the pGFP vector containing the Luciferase gene (hopefully the ligation worked this time).17:22, 15 May 2006 (EDT)
'''May 15''': Transformed Top10F' bacteria with the pGFP vector containing the Luciferase gene (hopefully the ligation worked this time).17:22, 15 May 2006 (EDT)
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'''May 16''': Completed agarose gel extraction of pGFP plasmid DNA, as cut by EcoRI and HindIII.
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'''May 16''': Transformation of Top10F did not work.  The cause was an error in the extraction of the DNA from the the agarose gel.  Completed a new agarose gel extraction of pGFP plasmid DNA, as cut by EcoRI and HindIII.  Prepared a new seed of Top10F bacteria transformed with luciferase.  Prepared a midiprep seed of EcoMscL.
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'''May 17''': The control of the miniprep of luciferase was murky.  This could have been due to contaminants in the LB, water, ampicillin, or pipet tips.  A miniprep of both luciferase and EcoMscL was conducted.  A new seed of luciferase was prepared.  The miniprep of luciferase was digested with EcoRI and HindIII.  The miniprep of EcoMscL was digested with XmnI.  A gel with the digests was run but the EcoMscL showed no bands and the luciferase only had faint bands.
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'''May 18''': The controls of the seed of luciferase were clear.  A miniprep of the new luciferase seed and the old EcoMscL seed was conducted.  The same restriction enzymes were used to digest the plasmids.  A gel was run with the digests.

Revision as of 19:40, 18 May 2006

May 15: Transformed Top10F' bacteria with the pGFP vector containing the Luciferase gene (hopefully the ligation worked this time).17:22, 15 May 2006 (EDT)

May 16: Transformation of Top10F did not work. The cause was an error in the extraction of the DNA from the the agarose gel. Completed a new agarose gel extraction of pGFP plasmid DNA, as cut by EcoRI and HindIII. Prepared a new seed of Top10F bacteria transformed with luciferase. Prepared a midiprep seed of EcoMscL.

May 17: The control of the miniprep of luciferase was murky. This could have been due to contaminants in the LB, water, ampicillin, or pipet tips. A miniprep of both luciferase and EcoMscL was conducted. A new seed of luciferase was prepared. The miniprep of luciferase was digested with EcoRI and HindIII. The miniprep of EcoMscL was digested with XmnI. A gel with the digests was run but the EcoMscL showed no bands and the luciferase only had faint bands.

May 18: The controls of the seed of luciferase were clear. A miniprep of the new luciferase seed and the old EcoMscL seed was conducted. The same restriction enzymes were used to digest the plasmids. A gel was run with the digests.

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