Pharmacia genomic DNA kit

Dietmar Baas uc05 at rzstud1.rz.uni-karlsruhe.de
Thu Feb 23 16:21:25 EST 1995


TUCKER at AFP76.HUMC.EDU wrote:
: Netters,
: 	Has anybody used the Pharmacia (or any other) genomic DNA preparation 
: kits?  
: Do they work?  Are they as fast and easy as advertised?  Any other coments?
: Thanks in advance for your help.
: 		Mark

Hi, Mark

I use the QIAGEN Genomic-tip 100/G (MIDI Prep, up to 100 ug DNA) for 
isolation of gen. DNA from Gram-neg. bacteria. From 4 ml over night culture
of E. coli I got 70 ug pure DNA, no RNA contamination (on an agarose-gel),
the A260/A280 ratio is about 1.8 and the length of the DNA between 50 to 
100 kbp.
The handbook contains protocolls for isolation from bacteria, yeast, tissue,
blood, cultured cells.
The procedure needs less than 3 h from harvesting the cells to 
iPrOH-precipitation.
The price for 25 tips without buffers (I've prepared them by myself, you
need only NaCl, GuHCl, MOPS, Tris, RNase, Prot K, Lysozym (for bacteria))
is 220 DM (perhaps about 150 $).
Compared with the protocolls from the "Maniatiss" (I've tested Protocol 1 &
3, but always huge amounts of RNA) the kit works fine, and I think so do the
kits from other companies.

At last, a little advice for better yield: elute with prewarmed buffer 
(50'C) and delay the elution by locking the tip for 30-60 min (I put
the tips in an 37'C oven during this time).
This works also for all other column based isolation methods, i.e. plasmid
isolation. 

Greetings from rainy germany, 

Dietmar




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