Unspecific hybridization to plasmid-vector

Manuel G. CLAROS claros at obelix.cica.es
Mon Mar 4 14:29:12 EST 1996


Se me ponen los ojos a cuadros cuando David Grant nos cuenta:

 > > Hybridization of my probe to the vector is weaker than to a true positive,
 > > but quite strong, so that I can see clear bands after 2h exposition.
...
 > phenol extraction - the DNA was then run again on a 1% gel, the bands cut
 > out and DNA purified and run AGAIN on a 1% gel - this gel was blotted and
...
 > I have no good explanation for this but have seen it for years

DNA bands in agarose are contaminated with DNA from other bands. Try tu
PURIFY TWICE your probe and you will diminish dramatically the plasmid
hybridztion

-- 
Manuel G. CLAROS     -. .-.   .-. .-.   .   -> claros at obelix.cica.es <-
Biologia Molecular   ||X|||\ /|||X|||\ /|   No quiero cambiar, no puedo,
Facultad Ciencias    |/ \|||X|||/ \|||X||   no quiero ser una frase mas.
E-29071 Malaga       '   `-' `-'   `-' `-         (Esclarecidos)



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