hybond

Edwin you at somehost.somedomain
Tue Jan 21 05:14:01 EST 1997


In article <mikechao-ya023080001601971440010001 at news.swmed.edu>, mikechao at hamon.swmed.edu (Mike Chao) says:
>
>In article <33EC032025 at puskin.sote.hu>, SZABADKAI at PUSKIN.SOTE.HU
>("Szabadkai Gyorgy") wrote:
>
>>Hi,
>>
>>I tried to use Hybond N membranes for dot blots of RNA and DNA, but 
>>the DNA signal has lost somehow. I denaturated the DNA by boiling for 
>>5 minutes and then fixed on the membrane by UV crosslinking (1200 
>>mJ/cm2). The probe was ok, I checked it.
>>What kind of denaturation and fixation do you use for this memebrane?
>>
>>
>>Szabadkai Gyorgy
>
>for denaturing we use akaline (i.e., the standard NaOH/NaCl solution for
>most Southerns), UV X-linking for fixation. For Hybond we found that if you
>wash the membrane with clean H2O after X-linking and just before adding hyb
>buffer, this reduces background.
>
>Mike
>
>--------------------------------
>Mike Chao                                       
>UTSW Medical Center @ Dallas
>Dept. Mol. Biol. & Oncology
>Phone: (214) 648-1489
>Fax:   (214) 648-1488
>http://hamon.swmed.edu/~mikechao
>--------------------------------


Hi, I heard the hybond membranes have been changed. this has caused us 
already some background problems on chemiluminiscention protocols. The background was 
very high making specific probing inpossible.


Edwin.





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