Team:Paris/July 25

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Contents

Extraction of the DNA

  • 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))
  • Test of the succeed of the extraction by electrophoresis on 2µl of the parts extracted.


==> 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

MiniPreps / Digestion

  • Minipreps : Protocol of Promega has been done on all the clones cultivated on the 24th.


Digestion Mix

10µl of Miniprep
12.5µl of water
2.5µl of Buffer N°2
0.25µl of BSA 100x
1µl of enzyme 1
1µl of enzyme 2


list of digestion

Name Miniprep Biobrick Enzyme 1 Enzyme 2 Nber tube
D100 MP100 B0034 XbaI PstI 2
EcoRI XbaI 1
SpeI PstI 2
D116 MP116 J23100 SpeI PstI 2
D120 MP120 B0030 SpeI PstI 2
EcoRI XbaI 1
XbaI PstI 2
D121 MP121 E0422 XbaI PstI 1 !
D122 MP122 E0840 XbaI PstI 1 !


Results of digestions : Electrophoresis

Name BioBrick Tube N° Enz 1 Enz 2 Obs Exp Size Matrix Exp Size BB Mea Size Matrix Mea Size BB
D100 B0034 1 XbaI PstI BI 2057 pb 34 pb
2
3 EcoRI XbaI FV 2076 pb 15 pb
4 SpeI PstI BV 2077 pb 14 pb
5
D116 J23100 1 SpeI PstI BV 2100 pb 883 pb
D120 B0030 1 XbaI PstI BI 2057 pb 37 pb
2
3 EcoRI XbaI FV 2079 pb 15 pb
4 SpeI PstI BV 2080 pb 14 pb
5
D121 E0422 1 XbaI PstI FV 2057 pb 939 pb
D122 E0840 1 XbaI PstI FV 2057 pb 900 pb
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