Extracting Plasmid DNA by the Alkaline Lysis Method (BIOL310)
Channel: It's Learnable
Date: 2020-09-07

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