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:
>>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 Chao                                       
>UTSW Medical Center @ Dallas
>Dept. Mol. Biol. & Oncology
>Phone: (214) 648-1489
>Fax:   (214) 648-1488

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.


More information about the Methods mailing list