DNA from FORMALIN fixed SPECIMINES

Will Fischer wfischer at bio.indiana.edu
Fri May 5 11:54:14 EST 1995


Guy Reeves (R.Guy.Reeves at ncl.ac.uk) wrote:
: Hi 
: I am about to attempt to amplify DNA from 
: formalin (= formaldehyde) preserved biological 
: specimens ...
: I am starting form the premise that ... the 
: difficulties in 200bp+ amplifications result form 
: the formation of formalin catalysed protein 
: DNA cross-linking...

Formaldehyde has been used for DNA-protein cross-linking because it is
reversible (so claim these authors):

Orlando, V. and Paro, R. (1993)  Mapping polycomb-repressed domains in
the bithorax complex using in vivo formaldehyde cross-linked chromatin.
Cell 75:1187-1198 (Dec 17 1993).

They got the cross-links off by SDS-proteinase K treatment... but I
suspect that the cross-linking they dealt with wasn't as extensive as
you might find in preserved tissue.  You could also check out the
following:

Solomon, M.J. and Varshavsky, A. (1985)  Formaldehyde-mediated
DNA-protein crosslinking: a probe for in vivo chromatin structures.
Proc. Natl. Acad. Sci. USA 82:6470-6474.


-- Good luck.
____________________________________________________________
Will Fischer				wfischer at indiana.edu

Department of Biology                   Lab:    812-855-2549
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Indiana University                      FAX:    812-855-6705
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