Team:Warsaw/Calendar-Main/11 July 2009

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Gel out phoP/phoQ

Kama

  • Isolation of fragment of the correct lenght(˜2200bp)from gel was performed with the A&A "Gel-out" kit.
  • construct BBa_K177012- operon1 part2

    Contents

    Ania

    Task1:

    Competent cells were transformed with BBa_C0012 - lacI repressor taken out of the distribution 200 Kit Plate 1 well 2O. It is on the pSB1A2 ampicillin resistant plasmid.

    Task 2:

    Alkaline lysis of bacterial cultures to obtain plasmids containing following bricks: BBa_R0051 - promoter (lambda cI regulated) BBa_B0032 - RBS.3 (medium)

    Task3:

    Digestion to confirm plasmid extraction. DNA that should contain R0051 on pSB1A2 plasmid was digested with PvuI and HindII. Digestion mix contained 5ul of extracted DNA, 2ul of Fermentas BamHI buffer , 0.3ul of each enzyme and water added to obtain 20ul total volume.

    The expexted fragments are: 695bp and 1447bp.

    April
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    October
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