Template:Team:KULeuven/13 August 2009/VanillinProduction

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(New page: * Miniprep on Sam8 and Ech *)
 
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* Miniprep on Sam8 and Ech
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# Plated ''COMT'' (+RBS) from the registry
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*
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#* Growing overnight in 37°C
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# Plated ''ech''
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#* From newly made liquid culture
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# Miniprep of ''ech'' and ''sam8''
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#* Nanodrop results:
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{| border ="1" align="center"
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!| Part || concentration (ng/μl) || 260/280 λ || 260/230 λ
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|- align="center"
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| ''ech'' ||72,7 || 1,94 ||
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|- align="center"
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| ''sam8'' ||66,2|| 2,00 || 
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|-
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|}
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# Restriction digest on ''ech'' and ''sam8''
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# Gel electrophoresis on ''ech'' and ''sam8''
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# New electrocompetent cells were used for the electroporation with the old ligations
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#* Lig A -> ''sam5 + sam8''
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#* Lig C -> ''ech + fcs''
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#* Note: there was no spark
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# Plating of the electroporated cells
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#* 200 μl per plate twice
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#* 600 μl left of A and C

Latest revision as of 10:01, 14 September 2009

  1. Plated COMT (+RBS) from the registry
    • Growing overnight in 37°C
  2. Plated ech
    • From newly made liquid culture
  3. Miniprep of ech and sam8
    • Nanodrop results:
Part concentration (ng/μl) 260/280 λ 260/230 λ
ech 72,7 1,94
sam8 66,2 2,00
  1. Restriction digest on ech and sam8
  2. Gel electrophoresis on ech and sam8
  3. New electrocompetent cells were used for the electroporation with the old ligations
    • Lig A -> sam5 + sam8
    • Lig C -> ech + fcs
    • Note: there was no spark
  4. Plating of the electroporated cells
    • 200 μl per plate twice
    • 600 μl left of A and C