User:Hetmann/Notebook/2006-11-7: Difference between revisions

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Latest revision as of 17:09, 14 November 2006



Goals for November/December

  1. Relabel and organize tubes within BL5088 and BL5096.
  2. Starting the week of Monday, 11/13, we are shooting for a goal of one successful ligation per week. Ligations of all constructs should be complete before Christmas break.
    • Ligations:
      1. 'J04500+KaiA+J04500+KaiC'+'J04500+KaiB'
      2. 'Promoter(rbs)+'KaiA'
      3. 'B0015'+'J04500+KaiB'
      4. 'B0015'+'J04500+KaiC'
      5. 'B0015+J04500+KaiB'+'B0015+J04500+KaiC'
      6. 'Promoter(rbs)+KaiA'+'B0015+J04500+KaiB+B0015+J04500+KaiC'
    • Progression
      • Step 1: Midiprep
        1. KaiA
        2. B0015
        3. J04500+KaiB
        4. J04500+KaiC
        5. Promoter(rbs)
        6. J04500+KaiA+J04500+KaiC
      • Step 2: Digestion
        1. KaiA/XP
        2. Promoter(rbs)/SP
        3. B0015/SP important
        4. J04500+KaiB/XP important
        5. J04500+KaiC/XP
        6. J04500+KaiA+J04500+KaiC/SP
      • Step 3: CIP/Gel Purification
      • Step 4: Ligation/Transformation
        1. 'J04500+KaiA+J04500+KaiC'+'J04500+KaiB'
        2. 'Promoter(rbs)+'KaiA'
        3. 'B0015'+'J04500+KaiB'
        4. 'B0015'+'J04500+KaiC'
      • Step 5: Midiprep
        1. Promoter(rbs)+KaiA
        2. B0015+J04500+KaiB
      • Step 6: Digestion
        1. Promoter(rbs)+KaiA/SP
        2. B0015+J04500+KaiB/XP
      • Step 7: CIP/Gel Purification
      • Step 8: Ligation/Transformation
        1. 'Promoter(rbs)+KaiA'+'B0015+J04500+KaiB'
      • Step 9: Midiprep
        1. Promoter(rbs)+KaiA+B0015+J04500+KaiB
        2. B0015+J04500+KaiC
      • Step 10: Digestion
        1. Promoter(rbs)+KaiA+B0015+J04500+KaiB/SP
        2. B0015+J04500+KaiC/XP
      • Step 11: CIP/Gel Purification
      • Step 12: Ligation/Transformation
        1. 'Promoter(rbs)+KaiA+B0015+J04500+KaiB'+'B0015+J04500+KaiC'
  3. Once J04500+KaiA+J04500+KaiC+J04500+KaiB is complete, borrow a chemostat (or build one? =\).
    • Test the construct, using Western Blot analysis.
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