As promised, this is the method I currently use for making maxiprep DNA.
I learned this method from an American in the lab where I did my DPhil. At the time, if I wanted to transfect eukaryotic cells I had to perform a double caesium chloride prep based on isopycnic ultracentrifugation.
That was as completely horrible as it sounds.
Marc introduced me to this method. It uses phenol, which is a bit scary (if you know what you're doing the risk is minimal), but produces superb quality plasmid DNA that can be transfected into cells with no further purification. And naturally you can use it for digests, transformation of bacteria, etc. It's cheaper than any commercial kit, probably faster, and certainly more robust. For the multi-tasking (or just plain lazy. . .) scientist, it is full of breakpoints where you can leave the prep and go and do something else, rather than being tied to the bench watching a column drip.
I couldn't do my job without it. Thanks, Marc.
Comments
Thanks! It looks great. I have something similar for a boiling minipreps but much, much (quicker and) dirtier - this looks spanking clean. If I ever have to go back to using preps again, I'll definitely go this way. I've used glycogen in RNA precipitations; perhaps I could use PEG 8000?
And phenol is way less scary than CsCl gradients. Brrrrr.
By the way, don't get all hot and bothered that you can't read my blog, neither can I. Vacation, sort of, thanks to the evil forces of the Internet.
Posted by: Alethea | February 13, 2008 01:34 AM
Ew. boiling minipreps? Fish out blob with toothpick. No thanks - I'll post my method.
Noticed the daemons had escaped from scienceboard.net. Hope you recapture them presently.
Posted by: bk | February 13, 2008 09:50 AM
Yes, but I like the home-cooked aspect of going to the local supermarket and buying toothpicks to autoclave. But I look forward to seeing your method. And I appreciated your note that reagents don't need to be fresh, I have used the same solutions for many years without ill effects.
Posted by: Alethea | February 14, 2008 06:18 AM