Team:Paris Liliane Bettencourt/Notebook/2010/07/22/

From 2010.igem.org



Antoine


Résults transformation biobrick R0040/C0040/R0011/C0012

Positif for all


Minipreps

pBM43 From 10 ml of overnight culture to 60μL Vf.


electrophoresis Minipreps

5μL of each in wells.

3μL of 1kb Ladder

picture of the gel



Overnight culture (biobrick R0040/C0040/R0011/C0012 in LB + Amp)*2 (one for glycerol)

  • P1003 LB+Amp+Kan
  • YejF KO (from KEIO collection , KanR disrupt YejF gene) (LB+ Kan)*2 (one for glycerol)

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Aleksandra, Stéphane, Raphaël
Minipreps : 60 minipreps in total, 4 minipreps of a 5ml culture per biobrick, eluted with 30μL buffer and collected into 1 tube for a total volume of around 120μL:

  • I732006 (LacZ)
  • B0031 (weak RBS)
  • B0032 (medium RBS)
  • B0034 (standard RBS)
  • J31007 (TetA C)
  • C0062 (LuxR w/o RBS)
  • J37033 (RBS/LuxR) from the 2007 distribution plate
  • 23119 (Ps)
  • J23110 (Pm)
  • attC
  • term/attC/mRFP
  • B0015 (term)
  • E0240 (RBS/GFP/term)
  • K081008 (LuxI)
  • R0062 (pLux)


Aleksandra, Raphaël, Théotime
Nanodrop to determine the amount of DNA in the miniprep samples:

Sample name 260/280 260/230 ng/μL
term (B0015)1.82 2.34 119.9
attC1.72 2.21 26.8
RBS/LuxI (K081008) 1.85 2.35 102.3
Pmed (J23110) 1.872.34 194.2
Pstr (J23119) 1.86 2.31 169.5
LuxR (C0062) 1.862.41 188.0
TetA(C) (J31007) 1.86 2.35149.1
RBS/LuxR (J37033)1.81 2.28 114.4
stand.RBS (B0034)1.86 2.30 145.2
med.RBS (B0032)1.90 2.34 120.1
weakRBS (B0031) 1.862.32 163.2
LacZalpha (I732006) 1.86 2.42 221.3
pLux (R0062) 1.93 2.27 66.6
t/attC/mRFP 1.97 2.13 33.1
RBS/GFP/term (E0240)1.892.39120.6