Best plasmid preps for Big-Dye sequencing

Susanne Rohrer srohrerNOSPAM at immv.unizh.ch
Tue Nov 30 10:51:32 EST 1999


Student wrote:

> Can anyone recommend which is the best plasmid miniprep kit for the
Big-Dye
> Sequencing reaction? I would prefer kits that provide eluted plasmid
which
> can go straight into sequencing without performing EtOH pptn first. I
've
> tried Hybaid (not so good), Invitrogen SNAP (a bit better but still
not
> ideal) so far but they haven't been too satisfactory and a lot of low
> signals (when I am lucky) or no signals (most of the time) at all!

"I just use 1/10 vol. NaAc.(3M pH round 5) and 2.5 vol. EtOH (100%)
spin max. speed 10-20 min
wash with EtOH (70%)"

I don't think that was what you expected. Try Macherey-Nagel or Quiagen,
they elute with a dilute Tris buffer, and come out quite clean.





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