Team:Warsaw/Calendar-Main/23 August 2009

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Assembly of endosomal detection operon

Marcin


Task 1:

  • Transformation of chemocompetent E. coli strain DH5&alpha

Constructs to transform:

Methods:

  • Detailed protocol of transformation is described here.

Task 2:

  • Isolate of plasmids from bacterial cultures prepared in 22.08.09

Methods:

  • Plasmids were isolated using the A&A plasmid mini kit. Detailed procedure of the isolation is described here

Task 3:

  • Digest previously isolated plasmids and verify the correctness of the ligation:

Methods:

  • Reaction mixture composition:
  •  
    2 μl purified plasmid DNA product
    0.5 μl XbaI (Fermentas)
    0.5 μl PstI (Fermentas)
    2 μl Buffer Tango (Fermentas)
    15 μl MQ water
  • Digest was performed one hour
  • Results:

    All isolated samples have expected plasmid

    Making of the plac-RBS-llo-intA part

    Jarek


    Tasks:

    • Transformating the competent ''E.coli'' strain TopF' with ligation
    • Plating the transformated culure on LB+Amp plates

    Cloning switch 1 regulatory parts [ K177012-PcI.RBS.LacI, K177033-PcI.RBS.LacI.PcI.RBS.RFP.terminator, K177011-PLacI.RBS.cI.terminator, K177038-PLacI.RBS.cI.terminator.PLacI.RBS.GFP.terminator ] into two compatible low copy number plasmids of different antibiotic resistance

    Ania


    Tasks:



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