Best plasmid preps for Big-Dye sequencing
jelitto at lion-ag.de
Mon Nov 29 12:13:32 EST 1999
> 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!
> Any comments will be appreciated. Thanks.
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%)
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