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    7/11: Miniprep and Sequencing of AmilCP, tsPurple, pSB1A3, and mRFP in pSB1K3. Plus parafilming
    • Notes
    • Metadata
    • Relevant Items
    • Measuring dsDNA concentration using the Nanodrop
    • DNA Fragment sample preparation for Genewiz Sanger Sequencing
    • Miniprep Plasmid DNA Purification from E. coli
    Editing disabled on read-only entry.

     
    7/11: Miniprep and Sequencing of AmilCP, tsPurple, pSB1A3, and mRFP in pSB1K3. Plus parafilming

    Thursday, 7/11/2019ObjectiveAttendanceMiscellaneousThings BoughtMiniprepsNotesMiniprep ResultsSequencingPrimer Preparation of VF2 and VRSequencingParafilming
    Thursday, 7/11/2019
    Objective
    Miniprep the following:
    ○
    amilCP BBa_K592009
    ○
    pSB1A3 pSB1A3
    ○
    pSB1K3+mRFP BBa J04450
    ○
    tsPurple BBa K1033906
    Sequence the following:
    ●
    amilCP BBa_K592009
    ●
    tsPurple BBa K1033906
    Attendance
    Sonia & Liana
    Miscellaneous
    Threw out 7/9 SOC due to contamination. We still have another 50mL tube of SOC, though
    Parafilmed the following plates and stored in Chilly:
    ●
    C.7.9.F through C.7.9.I
    Things Bought
    p20 LTS tips $3
    1.5mL microcentrifuge tubes $2
    Minipreps
    Notes
    M.7.11.B and M.7.11.C are stored in the -20 fridge iGem box: F2 and F3
    M.7.11.A and M.7.11.D are stored in -20 fridge iGem box: E5 and E6
    Overnight used for miniprep was from the following transformations:
    Sequencing
    Notes
    ●
    For Genewiz's purposes, our miniprep conenctrations have to be above 50ng/uL for Sanger Sequencing
    ●
    Primers must be diluted to 5uM with 5uL of primer sample per reaction.
    ●
    We are NOT sequencing pSB1A3 and pSB1K3. The reasons:
    ○
    Sequencing only good for about 800bps from primer before it becomes wonky, so we'd only get 1600 bps or so of good sequencing before it gets weird and that doesn't cover the whole plasmid (like 2100bps)
    ○
    Because of our plates, we know the amp/kan resistance gene works
    Parafilming

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