Template:Team:KULeuven/13 August 2009/VanillinProduction

From 2009.igem.org

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