Probing native DNA on filters(??)

GIDEON S. GORDON ggordon at opal.tufts.edu
Fri Sep 18 18:18:00 EST 1992


Greetings Netters!

    	My question may be simple, but we don't seem to have
anyone here who has attempted this.  Has anyone had any experience 
with blotting native DNA (ie.  Chromosome digest, non-denatured) 
onto nylon filters and subsequently probing this DNA with a DNA
binding protein?  I understand it is simple to blot native DNA
but will UV crosslinking fix it to the filter without having 
previously denatured it?
	One would assume that a DNA binding protein would require
the DNA binding site to be 1) in native form and 2) free from
binding to the filter.  It would be nice if this method could 
work and be used with a chromosomal digest blot or (even better)
a lambda bacterial library via colony hybridization.
	If anyone has any information, it would be greatly 
appreciated.  
			Thanks,
				G. Scott Gordon
				Tufts U. - Sackler School



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