wizard dna clean up

d.a.bailey gen124 at nof.abdn.ac.uk
Tue Nov 29 14:40:09 EST 1994


Michael Cooley (szcooley at chip.ucdavis.edu) wrote:
: PBS::NELSONJ (@PBS.DFO.CA) wrote:
: : Hi,
: : 	I've recently switched to using the wizard dna clean up system
: : for purifying fragments from lmp agarose gels.  Typically I Etoh precip.
: : the dna to concentrate it after eluting from the column.  I have been
: : having problems getting the dna into solution after this precipitation.
: : Has anyone else had this problem or noticed any particularities after
: : purifying dna from lmp agarose using the wizard system?
: : John Nelson
: : nelsonj at pbs.dfo.ca


: Are you sure its the DNA that wouldn't go into solution? Promega has been 
: going through periods with these columns. Some of them leak the resin. I 
: suggest you ignor the pellet and check the super on a gel.


: #%#%#%#%#%#%#%#%#%#%#%#%#%#%#%#%#%#%#%#%#%#%#%#%#%#%#%#%#%
: %                                                        #
: #   Dr. Michael Cooley         Practice Random Kindness  %
: %   mbcooley at ucdavis.edu    and Senseless Acts of Beauty #
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Just thought I'd add my 2p's worth, Michael Cooley is right it's more likely to
be the resin leeking through the column thats in the bottom of your tube.

	I've just switched to the Promega kit and found it works really well,
the recovery rates are pretty good even when you elute in a smaller volume -i.e.
20ul.  Which may save you the bother of a ppt step.  Be careful not to disturb
the resin left in your tube when you take the elutant out it seems to inhibit
ligase (at least thats my excuse ;-) 

	Another thing is the isopropanol wash, if you are not using a
vaccuthingy I think the protocol reccomends a 20sec spin to dry the sample
before elution. I found that was not long enough and there was still some
isopropanol coming through with the elution step. 
--

______________________________________________________________________
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              | Molecular and Cell Biology, |  
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              | d.a.bailey at abdn.ac.uk       |
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