UV-fixing of filters.
bhjelle at vaxine.unm.edu
Tue Dec 17 17:34:04 EST 1991
In article <9112161919.AA22055 at alize.ERE.UMontreal.CA> coady at ERE.UMontreal.CA (Coady Michael) writes:
>Dear Fellow Netlanders,
> Has anyone experience with UV fixation of nucleic acids
>to filters? Yes, I've used Stratalinkers before but I don't believe
>that I have access to one here. What I do have access to is a
>transilluminator and I spoke to someone at Boehringer Mannheim this
>morning who assured me that I should put my charged nylon on this,
>(face down, natch :-)) and leave it for five minutes. Yet this is
>quit a long time right next to a strong UV source, and I can't help
>but wonder if the nucleic acids are going to badly degraded by this
>method. Thanks in advance for any advice.
I've been doing this for years for everything from Southerns to
PCR dotblots. I use 5' on a 302nm transilluminator with good
results. If you are worried about 5' being too long, you can
try 2'. For PCR dotblots the people in the lab next to mine often
leave the box on for more like 20' then amble back to it when
they remember. Seems to work fine.
Incidently, don't do this with nitrocellulose, only nylon.
I'm putting on a flame-proof vest now:
Stratalinkers are a useless ripoff if you have access to a UV box.
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