pcDNA3 with Wizard

[Yasha Hartberg]

In article <1994Oct3.162722.11067 at midway.uchicago.edu>,
lshekter at kimbark.uchicago.edu (lee shekter) wrote:


> 
> I've used the Wizard system on pcDNA3 and haven't had any problem with res.
> enzyme cutting save for EcoRI, which doesn't appear to cut any DNA isolated by
> the Wizard method.
> -- 

I've never had any problems cutting with EcoRI on pBR322 derivatives after
Wizard Minipreps.  However, since Promega switched from the good ol' Magic
Miniprep, I can find no good reason to use the kits.  The yield is usually
very low, the quality is inconsistent, and yes, sometimes you don't get
good cutting with restriction enzymes.  All of this, Promega claims, was
necessary because the proprietary resin in the Magic Minipreps was too
inconsistent from their suppliers (read, since no one was having any
problems before, the suppliers obviously raised their prices and Promega
didn't want to pay).

Anyway, I suggest give up on the Wizard Minipreps!  Go back to the basics
such as alkaline lysis or boiling lysis preps.  With all of the steps you
have to add to clean up Promega's Preps, you end up saving time and money,
and generally get better, cleaner yield with some of these other methods.

Yasha Hartberg
Texas A&M University
"If people want to spend their whole lives creaming and tweezing and
brushing and tilting and gluing, that's really okay too, because it gives
them something to do."  Andy Warhol