Team:Paris-Saclay/Project/Notebook/Week 10

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====6th August====
====6th August====
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Revision as of 21:01, 25 September 2012

GEMOTE

GEMOTE

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6th August

  • Sending of the B1 sample to sequencing with two pairs of primers
    • K-274100 Forward and Reverse
    • Plasmid pSB1A2 Forward and Reverse


7th August

  • Liquid culture of B1 in order to prepare a glycerol stock
    • Receipt of new primers
    • 2 Forward
    • 3 Reverse
    • Plasmid Reverse


8th August

Amplification of the plasmid pSB1A2, BBa_K274100 and BBa_K115017 by PCR with the new primers. Visualization by electrophoresis on a 2% Agarose gel for BBa_K115017 and a 0.8% Agarose gel for BBa_K274100 and the plasmid pSB1A2. We are expecting a band at 147 bp for BBa_K115017, 3408 bp for BBa_K274100 and 2079 for the plasmid.
Week10-2.jpg


PCR program used: Week10-3.jpg

9th August

New amplification by PCR of the plasmid pSB1A2, BBa_K274100 and BBa_K115017 as it has been done the day before.
Week10-4.jpg
  • Digestion by EcoRI of the plasmid that contains BBa_K274100.


10th August

Amplification by PCR of BBa_K274100 already digested by EcoRI. Visualization by electrophoresis on a 0.8% Agarose gel.
Week10-5.jpg


PCR program used: Week10-8.jpg

Miniprep of BBa_K274100 followed by Nanovue to determine the concentration of the sample
Week10-6.jpg
Miniprep of the Plasmid pSB1A2 followed by Nanovue to determine the concentration of the sample
Week10-7.jpg
  • Digestion by HindIII of the plasmid pSB1A2 in order to linearize it.
Digestion of BBa_K115017 by DPNI to eliminate the plasmid matrix. Visualization by electrophoresis on a 2% Agarose gel. We are expecting a band at 123 bp.
Week10-1.jpg

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