I support the method suggested by John. I have used it several times
reproducibly. However, Rui, if you have to have a quick plasmid-prep protocol,
try the 2 minute "Insta MiniPrep Kit" by 5 Prime 3 Prime. Their address is :
5 Prime 3 Prime, Inc.
5603 Arapahoe Ave.
Boulder, CO 80303
Tel. 1-800-533-5703 (From U.S. & Canada)
or (303) 440-3705
Fax (303) 440-0835
Rajendra V. Deshpande
Pharmacia & Upjohn, Inc.
______________________________ Reply Separator _________________________________
Subject: Re: Need fast mini-prep protocols
Author: jrt at intac.com (John R. Thompson) at INTERNET
Date: 11/11/95 7:21 PM
lur at duke.usask.ca (Rui Lu) wrote:
>> Now I have a large number of colonies to screen for insert.
>Does anyone know any fast mini-prep procedures to yield enough, cut-
>able plasmid DNA?
>You could try a whole cell PCR approach. Use PCR with one primer
specific for the insert, the other primer specific for an adjacent
region on the plasmid. Alternatively, if the inserts are not too
large you can just use vector primers flanking the insert site too.
Now, just do whole cell PCR and run a gel to identify insert
John R. Thompson
Merck Research Labs