DIG fun, pressure sensitive membranes..?

Andre Hamel hamel at ccu.umanitoba.ca
Thu May 20 09:23:17 EST 1993


>>I have an `interesting question' for all those who have used the Boehringer 
>>Mannheim DIG DNA labeling and detection kit.
>
>>yesterday, I completed the final step (color reaction step) on two nylon 
>>membranes. On both membranes, there was clearly visible the text which I had 
>>written on the outside of the ziplock bags in which the prehyb/hyp took 
>>place.
>
>>I am curious: Does this indicate extreme pressure sensitivity of the 
>>membranes? (I don't press hard when I write with marking pens. The pen in 
>>question was a scripto med. point permanent black marker.)
>>Other option: Is this indicative of ziplock back permiability to the ink?
>
>>Has anyone had ANYTHING like this happen...?
>
>>Thanks!
>
>>........    Tim Chipman                ..................................
>>..........    901106c at axe.acadiau.ca       ..................................
>
>I use Lumiphos (the light-generating alk-phos. substrate) in my DIG-hybs, 
>and it doesn't even work with zip-lock bags.  All I have to do is touch the 
>filter-Ab-conjugate to a ziplock bag, and I get a black smear.  (I tried 
>this with half of a filter, and got half a smear, the other side was ok).
>
>I have to use the heat-seal bags that i do my hybs in for my exposures.
>
>
>cheers, John
>
>John Nash                           | Email: Nash at biologysx.lan.nrc.ca.
>Institute for Biological Sciences,  | National Research Council of Canada,
>Cell Physiology Group.              | Ottawa, Ontario, Canada.
>             *** Disclaimer:  All opinions are mine, not NRC's! ***

Hi John

(yeh .. I guess I do read ALL postings to this group ... try hard NOT to
reply to them all though  :-)

Have you tried using clear (not the ones with a frosted side) transparency
 sheets for chemiluminescent development? We use these (heat seal with
sealer that has WIDE (0.5 cm) heating bar instead of a wire .. otherwise
can leak ... we use CSPD (Tropix) instead of Lumiphos, due to lower price
and increased sensitivity. Never experienced background/fog, etc ...
always (so far) got developments similar to P32.

We had bear of a time finding clear heat seal bags, so in a pinch I went
to the stationary supply room, took one of many boxes of transparencies ...

regards
   ----------------------------------------------------------------------
Andre Hamel                                 email: hamel at ccu.umanitoba.ca
Manitoba Veterinary Services                lab tel.: (204) 945-7630
Infectious & Genetic Disease                FAX: (204) 945-8062
Mol.Biol.Lab                                
Winnipeg, Manitoba, CANADA
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