Team:Paris/July 25


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DNA Extraction

  • Use of Promega's protocol for the extraction of: gel n°1 --> 4 : (D100 --> D110 cl-1 ES))
  • Use of Qiagen's protocol for the extraction of: gel n°5 --> 8 : (D110 cl-2 ES --> D120))
  • Analysis of the extraction success by electrophoresis on 2µl of the parts extracted.

Determination of DNA concentration by spectrophotometry


  • 2µl of the DNA extracted / 98µl of pure water
  • Reading against the adaptated blank (2µl of the elution's buffer / 98µl of pure water)

Results :

-D100 --> D110 cl-1 ES : Absorbance ~20-50µg/ml ; DO 260/280 = 1,5 ; DO 260/230 = 0,0-0.20

-D110 cl-2 ES --> D120 : Absorbance ~0-10µg/ml

Determination of DNA concentration by electrophoresis

  • to check for the samples from Qiagen's protocol, if the absence of detection of absorbance, is due to a problem of reading or a problem during the extraction, we realise an electrophoresis.

==> conclusion : we don't succeed to detect DNA by spectrophometry or electrophoresis for the samples produced by Qiagen's protocol.

==> we decide to repeat all the samples from the beginning, so we culture O/N at 37°c all the strains usefull for the experiments


in 10ml LB with adaptated antibiotics, all the Biobricks used before (MP100 --> MP122)

  • will be use to do minipreps


  • Use of Promega's protocol on all the clones cultivated on the 24th.
  • Preparation of 50µl of minipreps in simple.

list of Minipreps

Name Biobrick Description
MP100 B0034 Strongest RBS (Efficiency = 1)
MP116 J23100 Strong constitutive promoter in J61002
MP120 B0030 Strong RBS (Efficiency = 0,6)
MP121 E0422 ECFP (RBS+LVA+Term)
MP122 E0840 gfp tri-part; strong rbs