Team:Paris/July 25

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
Conditions:


 * 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

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


 * will be use to do minipreps

MiniPreps

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