ssDNA (again? ...yes, again!)

Helge Weissig helgew at LJCRF.EDU
Tue Nov 2 21:48:25 EST 1993


Hi netters,

        this is a kind of follow-up to the ssDNA purification thread as
well as to my earlier question about the digestion of ssDNA via annealing
of an oligo.
Well, it seems that my problems were due to the purification procedure.
Although i did get reasonable sequencing results from my preps, i was not
able to cut that piece of s....sDNA. Now i have used a kit (booooooo) but
obviously w/ success (aaaaaaahhh) since the DNA is digestable (munch,
munch).
Has anybody any serious (huh mmmh) thoughts about that?

Regarding the electrophoresis of ssDNA i did use neutral gels now, since i
am still not very happy with the alkaline electrophoresis methods (50 or 30
mM NaOH, 1 mM EDTA as buffer, low voltage, many long hours, staining after
neutralization in 1.5 M NaCl, 0.5 M Tris-HCl pH 7.2, 1 mM EDTA w/ EtBr in
either TBE or TAE). i have seen really nice pictures of alkaline gels but
mine can only be used to scare little kids asking for Halloween candy
(boooh).

Oh, yeah, the name of the kit: QUIAGEN's M13 kit (low yield, but only one
prep done so far).

anything appreciated,
helge

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