Extracting Plasmid DNA by the Alkaline Lysis Method (BIOL310)

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  • เผยแพร่เมื่อ 6 ก.ย. 2020
  • In this video, I take you step-by-step through the entire procedure for a typical "plasmid mini prep". Throughout the video, I point out some specific things you should be doing and should be observing.
    00:32 - Preparation is important
    01:19 - What's in your ice bucket
    02:05 - The Phenol/Chloroform tube
    02:55 - Balancing the tubes in a microfuge
    03:55 - End of first centrifugation
    04:55 - Removing the LB from the pelleted cells
    06:45 - Vortexing your samples to resuspend the cell pellets (set the vortex to "Touch" and maximum speed)
    08:15 - Addition of Solution B and observations
    10:05 - Addition of Solution C and observations
    12:40 - Recommended orientation of epi tubes when centrifuging samples
    13:30 - Separation of supernatant from the pellet
    16:35 - Addition of Phenol/Chloroform (work quickly)
    19:20 - Separating the aqueous phase from the organic phase
    19:34 - Why am I wearing gloves?
    20:55 - Transferring out the aqueous phase
    24:45 - DNA precipitation
    28:20 - DNA pellets!
    31:09 - Ethanol wash
    33:05 - Drying your DNA pellets

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