Protocols for isolating large (>200kb) plasmids.

[Lynda]

In article <3487.2dee0b34 at mbcl.rutgers.edu> goyal at mbcl.rutgers.edu writes:
>From: goyal at mbcl.rutgers.edu
>Subject: Protocols for isolating large (>200kb) plasmids.
>Date: 2 Jun 94 20:39:16 GMT

>Dear netters,
>        I am looking for protocols for isolating 'mega' plasmids (200kb or more)
>from bacteria. I have tried some of the usual alkaline lysis procedures, but
>havent yet succeeded fully.
>        Before I test a couple of other procedures, I thought I would get input
>from others who might have had some experience in this.

>Thanks

>ANIL GOYAL

>Goyal at mbcl.rutgers.edu


At ASM this year, I saw a poster the described such a method, but I haven't 
tried it yet. Following in-gel lysis, you linearize the DNA with S1 nuclease, 
then run a pulse-field gel.  The 
poster number was H249, and the authors are Barton, Harding, and Zuccarelli 
from Loma Linda University.  The method hasn't yet been publised in a journal. 
 If anyone tries this method in the near future, I would be interested in 
knowing how it turns out.  Good luck!
-------------------------Everybody got mixed feelings----------------------
------------------------About the function and the form--------------------
--------------------Everybody got to elevate from the norm-----------------
                                            ---Rush
                                            ---(Not Limbaugh)

Lynda Callicotte
lcallico at utkvx.utk.edu