Plasmid purificaction

[R.C.Ogden]

Umnarj Paeratakul wrote:
> 
> Dear friend
> 
> I am looking for ways to purify big plasmid too.  Could you
> e-mail me the protocol if you get one?
> 
> Thanks in advance
> 
> Umnarj PaeratakulHi,
	This depends on what species you are dealing with and what you 
call "Big Plasmids". We work with "mega-plasmids/mini-chromosomes" from  
Pseudomonas which are 110 to 200+ kb. For this try the method of 
"Wheatcroft & Williams" it is in a Short Communication to Journal 
of Bacteriology (I think) around 1986. This works well but the DNA is not 
too stable so don't keep it for more than a week or so. For large 
plasmids in E.coli eg pKT231 clones around 26kb I use a fairly standard 
alkaline lysis miniprep. These yeild quite well if a host like XL1 Blue 
MRF' is used.

			Hope this helps					
						Richie

*************************************************************************
 Richard C. Ogden,               E-mail: r.c.ogden at bangor.ac.uk
 School of Biological Sciences,   Phone: 01248 351151 ext.2368
 (Memorial Building),             (International: +44 1248 351151 ext.2368) 
 University of Wales,               Fax: 01248 370731
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 LL57 2UW, UK.
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************************************************************
Richard C. Ogden,          Email: r.c.ogden at bangor.ac.uk
SBS, Memorial Building,     Tel : 01248 351151 ext.2368
University of Wales,              (+44 1248 351151 ext.2368)
BANGOR, GWYNEDD.            Fax : 01248 370731
LL57 2UW, UK.                     (+44 1248 370731)
************************************************************