UV crosslinking of DNA - wavelength?

Colin Rasmussen colin at pombe.usask.ca
Fri Aug 13 12:09:40 EST 1999


Jeremy Carson wrote:
> 
> I am planning to do hybridization of a labelled 25bp probe to 250bp PCR
> amplified DNA by dot-blot hybridization. I intend binding the amplified
> DNA to nylon membranes by UV crosslinking.
> My understanding is that the optimum wavelength for UV crosslinking of
> DNA to nylon membranes is 254nm. How effective is crosslinking at 312nm?
> If 312 nm is less effective than 254nm how does it compare to baking at
> 80C for 2 hours.
> Any comments?
> 
> Jeremy Carson

I don't know about 312nm.  With 254nm and a Stratalinker, 1200 J/m2
takes about 30 sec.

Colin Rasmussen
U of Saskatchewan



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