Team:Paris-Saclay/Project/Notebook/Week 11

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13th August

Purification by PCR clean-up of BBa_K115017 treated with DPNI. To visualize the fragment, an electrophoresis has been made with a 2% Agarose gel. We are expecting a band at the size of 123 bp.
Week11-1.jpg
Week11-2.jpg
New PCR of BBa_K115017 to obtain more quantity of the fragment


PCR program used: Week11-3.jpg

  • Digestion by DPNI of the BBa_K115017 in order to degrade the matrix plasmid.
Determining the concentration of the plasmid pSB1A2 which has been linearized by HINDIII. We are expecting a band at the size of (2079+935) ~3kbp
Week11-4.jpg
A new PCR of the plasmid pSB1A2 is realized to obtain more quantity of the plasmid. We are expecting a band at the size of 2079 bp.
Week11-5.jpg

PCR program used: Week11-6.jpg


14th August

Determining of the concentration of the BBa_K274100 by electrophoresis on a gel at O.8% Agarose. We are expecting a band at the size of 2100bp
Week11-7.jpg
  • Digestion of the BBa_K274100 by DPNI to degrade the matrix plasmid.
  • Stocking up cells with glycerol
    • PSB1A3Amil CP + Ampicilline
    • PSB1C3 Amil GFP + Chloramphinicol


15th August

Day of public holiday in France.


16th August

PCR of BBa_K274100 and visualization by 0.8% Agarose gel electrophoresis. We are expecting a band at 3400bp.
Week11-8.jpg

PCR program used: Week11-9.jpg

  • Digestion of the BBa_K274100 by DPNI to degrade the matrix plasmid.

17th august

Visualization of the DPNI’s digestion of the plasmid pSB1A2 by 0.8% Agarose gel electrophoresis. . We are expecting a band at the size of 2079 bp.
Week11-10.jpg
  • Miniprep of the plasmid pSB1A2.
  • Miniprep of BBa_K274100 and BBa_K115017
  • Gibson assembly of the B construction
  • Transformation of DH5α competent cells with the Gibson B construction. The Petri dishes are placed at 37°C.