Alas, poor "magic"! I knew thee well.

Warren Gallin wgallin at gpu.srv.ualberta.ca
Fri Mar 4 14:22:54 EST 1994


In Article <1994Mar2.152928.8680 at mbcf>, neale at mbcf.stjude.org wrote:
[stuff deleted]

>However, we use a very simple boiling method that processes minipreps in bulk
>using a microcentrifuge. These minipreps are crude but work well for sequencing
>or restriction analysis, and best of all, require no phenol or other "magic"
>reagents. The combination of plasmid and host cell that we use is pBluescript,
>and DH5 alpha or DH10B cells, respectively.
>
[stuff deleted]

Have you tried using the plasmids for dye terminator sequencing?  We've
found that this is much more sensitive to the way the template is prepared
than Sequenase with dideoxy terminators.  On the other hand, we reliably get
450 instead of 250 b.p. of seqeunce, so a little more cost and effort is
worth it.

I would love to hear of a quicker, cheaper way to make reliably high quality
template for Taq cycle sequencing with dye terminaters.  Any suggestions?

Warren Gallin,
Department of Zoology, University of Alberta
wgallin at gpu.srv.ualberta.ca



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