wizard DNA clean up
barrass at pplros.demon.co.uk
Thu Dec 1 10:37:27 EST 1994
In article <9411301742.AA16624 at fraser.sfu.ca> Richard Kenneth Bruce
Martel, richardk at sfu.ca writes:
>Howdy people. Has anyone used either the Wizard DNA cleanup or PCR
>preps to isolate a LARGE piece of DNA from a normal agarose gel.
>Their 'propaganda' says the systems should be able to handle pieces
>as large as 15 or 23kb. How have they worked in the field? What
>about Bio-Rad's Prep-a-gene? The piece I want to gel purify is a
>10kb fragment from a restriction digest.
We used to use Prepagene all the time for all frags of
sizes from 0.5 up to about 14 Kb. It works ok as far as size goes
(we were cloning and sometimes transfecting the DNA).
Although the frag sometimes looked a little fuzzy on a
gel (low salt or degradation?) I've never seen a smear below
it or anything loke that. But the yield from PAG is
awful IMH, especially with large scale preps (5-15%).
Currently I'm trying to use Wizard resin from the miniprep
kit (it was going to waste - honest). I've been melting the
agarose in NaI (PAG binding buffer, actually) and then
adding Wizard resin. The binding buffer precipitated once, but
washed away with column wash, and I got about 70% recovery and
the DNA worked fine in a ligation. What's in the Wizard clean up kit?
Is the resin the same as in the miniprep kit? Does it
More information about the Methods