genomic DNA contamination using Wizard plasmid kit

Neil Saunders neil.saunders at unsw.edu.au
Tue Jun 5 21:39:46 EST 2001


""Deanne Bell"" <dbell at qnis.net> wrote in message
news:200106051527.f55FRQp66026 at ns.qnis.net...
> I'm not quite clear - are you getting the restriction smear AND the band
in
> the same sample or do you get one or the other?

I'm not quite clear either!

When we run the sample without digesting, we get a discrete band at about
18-20 kbp.
If we do a "normal" digest (1 ul enzyme, 1 h), we see a smear reminiscent of
a chromosomal digest.
If we use less enzyme or less time, we see no digestion (this last one not
so relevant, presumably just need to "fine-tune" conditions.)

Basically, the digest makes me think of chromosomal, but the undigested
sample makes me think of large plasmid, hence our confusion.

Neil
--
School of Microbiology & Immunology,
University of New South Wales,
Sydney 2052,
NSW, Australia

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