Homemade FTA paper and kit???
(by bt8a002 from uni-hamburg.de)
Fri Jul 18 04:01:18 EST 2008
thanks a lot... how stupid of me not to come to the same conclusion..
> Nothing fancy there. It's just a thick filter paper with dried Tris,
> EDTA and SDS. You spot cell onto it, SDS lyses them, then
> the whole thing is dried. Quick wash with TE removes some
> DNA and most of the SDS/EDTA, then more DNA is eluted
> and used for transformation (when storing clones) or PCR.
> For cleaner things, phenol and/or isopropanol washes
> can be added before elution.
> It is described in the multitude of patents by inventor
> Leigh A. Burgoyne and Assignees either Flinders Technologies
> or Whatman. E.g., # 5496562:
> ~ 50 ul of 2% SDS, 10 mM EDTA, 60 mM tris spotted on
> 1 cm2 of Whatman 3MM and dried.
> FTA kits have some other filter paper, thicker than 3MM but
> it really does not matter. Also, the concentrations seem to
> be an overkill. Long ago, I soaked some 3 MM in just
> TE + 1% SDS, dried and spotted 5 ul of several overnight
> cultures. A year after, storing the paper in the drawer, all
> clones gave lots of transformants after electroporation.
> This is a good candidate for inclusion into lab generics Wiki
> found at http://methodsandreagents.pbwiki.com/
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