Team:UrbanTundra Edmonton/Notebook

URBAN TUNDRA ▤

Timeline

  • July 20th 2016

    1. 4 microlitres of milliq water with 1 microlitre of plasmid
    2. 5 microlitres of no chlorite dismutase
    3. 1 microlitre of cutsmart buffer (specifically 1.1111 microlitre)
        4. a. 10 x cutsmart buffer into tubes makes 1x [ ]
    4. When adding BsaI, only touch the surface of the BsaI to the pipette tip. The reason: has glycerol which is sticky. Take 0.5 microlitre of BsaI.
    5. Mix sample very gently then put into a 37 degrees celsius incubator for 1 hour.
    6. Heat kill at 65 degrees celsius for 20 minutes. This kills the BsaI to prevent star activity.
    7. Pulse down tubes then add 8 microlitre of milliq water.
    8. Add 2 microlitres of T4DNA ligase buffer.
    9. Add 0.5 microlitre of ligase then mix gently and incubate at room temperature for 30 minutes or longer

  • July 21st 2016

    1. Did some plating.
    2. Plated three agar types:
        3. A. 4 plates of no antibiotic
        B. 10 plates of kanamycin (CM)
        C. 10 plates of ampicillin (Amp)
    3. Purpose: try to find the antibiotic marker on bacteria.
    4. Observational expectations: If it’s purple, then there is no Cld gene sequence. If white then has CLd gene sequence.
    5. Lives or dies on certain plates. Alive on ampicillin, then it has the marker. Dies it has the CM marker.
    6. Added 20 microlitres in each plate
        7. A. A total of 7 plates
        B. DNA minus has none, CM, and AMP
        C. Positive control has CM and AMP
        D. Cld +/- has CM and AMP

    Competent cell transformation:

    1. 1 microlitre of positive control added to competent cells (positive control: CPB we used yesterday before cutting and gluing)
    2. 10 microlitre of cloned plasmid added to competent cells (Cloned plasmid is what we did yesterday).
    3. Positive control, negative control, and cloned plasmid were chilled for 30 minutes, heat shocked for 90 seconds, chilled on ice for 5 minutes, and finally incubated at 37 degrees celsius with 1 ml of LB broth.

  • July 22nd 2016

    + control DNA - Cld + Cld -
    No Antibiotic Yes, very thick
    Amp No growth No growth No growth No growth
    CM Growth (purple colonies) No growth Possibly purple and white* Possible purple and white*
    * Purple colour will develop more over larger period of time (perhaps over the weekend)

  • Mussum ipsum cacilds

    Mussum ipsum cacilds, vidis litro abertis. Consetis adipiscings elitis. Pra lá , depois divoltis porris, paradis. Paisis, filhis, espiritis santis. Mé faiz elementum girarzis, nisi eros vermeio, in elementis mé pra quem é amistosis quis leo. Manduma pindureta quium dia nois paga. Sapien in monti palavris qui num significa nadis i pareci latim. Interessantiss quisso pudia ce receita de bolis, mais bolis eu num gostis.

  • Mussum ipsum cacilds

    Mussum ipsum cacilds, vidis litro abertis. Consetis adipiscings elitis. Pra lá , depois divoltis porris, paradis. Paisis, filhis, espiritis santis. Mé faiz elementum girarzis, nisi eros vermeio, in elementis mé pra quem é amistosis quis leo. Manduma pindureta quium dia nois paga. Sapien in monti palavris qui num significa nadis i pareci latim. Interessantiss quisso pudia ce receita de bolis, mais bolis eu num gostis.

  • Mussum ipsum cacilds

    Mussum ipsum cacilds, vidis litro abertis. Consetis adipiscings elitis. Pra lá , depois divoltis porris, paradis. Paisis, filhis, espiritis santis. Mé faiz elementum girarzis, nisi eros vermeio, in elementis mé pra quem é amistosis quis leo. Manduma pindureta quium dia nois paga. Sapien in monti palavris qui num significa nadis i pareci latim. Interessantiss quisso pudia ce receita de bolis, mais bolis eu num gostis.

  • Mussum ipsum cacilds

    Mussum ipsum cacilds, vidis litro abertis. Consetis adipiscings elitis. Pra lá , depois divoltis porris, paradis. Paisis, filhis, espiritis santis. Mé faiz elementum girarzis, nisi eros vermeio, in elementis mé pra quem é amistosis quis leo. Manduma pindureta quium dia nois paga. Sapien in monti palavris qui num significa nadis i pareci latim. Interessantiss quisso pudia ce receita de bolis, mais bolis eu num gostis.