ds Sequencing Qiagen-prep Plasmids-Help!
landel at helios
Fri Aug 12 14:35:28 EST 1994
We can ds sequence plasmid DNA made by the STET boiling miniprep
DNA any time we want.
When we try the same protocol on DNA purified using a Qiagen maxi
prep, it doesn't work, ie, no bands at all. DNA that is too pure
to sequence seems a little strange. I thought maybe that it was
all supercoiled, so that it wouldn't be a good substrate for the
polymerase in vitro, but cutting the plasmid to linearize it gave
only the slightest improvement (I can now see bands, but they are
Does anybody have any ideas about how to fix this? It seems silly
to do a bunch of minipreps when in theory one big maxiprep should
give you a life time supply of DNA, but that is the fix we have been
reduced to using.
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