Nothern with 290bp probe

Gregor Bucher st2153 at zi.biologie.uni-muenchen.de
Tue Feb 22 05:43:48 EST 2000


>Hi all
>I would like to perform a Northern blot with a relatively short probe
>of 290bp. GC content is about 60%. Any experience with with similar
>probes under standard hyb conditions (65°)?

I hybridized a cDNA library with such a short probe. I lowered temperature
to 50 and washed with low stringency only. I used Hybond XL
Nylon-membrane. It is said that with short probes (<100) blocking reagent
should only be used in the PreHyb not in the Hyb buffer (because protein
seems to "quench" the signal, Maniatis 9.51)) 

Maybe some of these things do also aply to Northerns ?

Gregor
-- 
Gregor Bucher, Zoologisches Institut der LMU München
0049/89/5902/444
st2153 at zi.biologie.uni-muenchen.de
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