From 2014.igem.org

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24^th June

• Ligation pSB6A1 and VPGXG-20 of 23/06/14
  2µl of digestion product
  1,5µl insert - ratio 1 vector/3 insert
  1µl Buffer ligase 10X
  0,5µl T4 ligase
  H2O qps 10µl
  30 min at 16°C


• Digestion ELP-40 and -60 and vector pSB1C3

• Test preculture 200ml
  
  13h of incubation for 20 et 40 (Glycerol)
  13h of incubation for 60 (glucose)
  For the next preculture : 200ml / 13h of incubation / put less bacteria in the preculture medium.
  


• Seeding at 11h
  We want OD = 0,12 in 1L
  
  C1V1=C2V2
  V1 (preculture 20) = 0,12/2,777=43,2ml of preculture medium in 1L LB+Glycerol
  V (préculture 40) = 0,12/2,609=46ml -> 40/glycérol
  V (préculture 60) = 0,12/2,853=42ml -> 60/glycérol
  
  
  DO T=0 (11h10)
  20: 0,164
  40: 0,093
  60: 0,160
  
  T=9h30 (17h40)
  20: 1,802
  40: 1,713
  60: 1,508