Midiprep kit with microfuge only? Promega MagneSil?

Thomas Landers thomas at genome.wi.mit.edu
Wed Apr 5 15:35:46 EST 2000


Hi,
We use Qiagen columns for purifying BAC DNA for library construction.
These kits use an isopropanol precipitation to recover the DNA from the
elution buffer. 
We found that 15.000 x g wasn't necessary, we precipitate the DNA in a
bench top centrifuge (about 3500 x g) for the same time (30 min). We are
very happy with this, as it allows the use of conical bottom tubes. Try
it in the 50 mL (or 15 mL) falcon tubes.
Tom Landers

John Dixon wrote:
> 
> Hi, can anyone help me find a transfection quality midiprep (50-100ug
> DNA needed) kit that either doesn't need a final ethanol precipitation
> step, or has a DNA precipitation step in a small enough volume for
> spinning in a microfuge - in a single eppy preferably (we are fed up
> with splitting our DNA preps into eppys, as we don't have access to
> high speed large volume fuges. We do have a bench top fuge to spin the
> crud out at 4000rpm in 50ml falcons prior to precipitation so a prep
> requiring low speeds would also suffice).
> 
> The promega Wizard PureFection plasmid prep kit which binds the plasmid
> DNA with paramagnetic beads looks like it could be tweaked to give a
> small enough volume for single eppy precipitation, although it needs a
> lot of hands-on washes. Has anyone had experience with this or any
> other suitable kits/protocols?
> 
> Any advice appreciated (suppliers or affils, please feel free to e-mail
> me direct if you have what I need, to avoid irritating the newsgroup
> with adverts!)
> 
> Cheers
> JD
> 
> --
> John Dixon                    Lab 44 (1223) 334131
> Wellcome/CRC Institute        Fax 44 (1223) 334089
> Cambridge University
> United Kingdom CB2 1QR       e-m: jpcd100 at mole.bio.cam.ac.uk




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