in situ hybridization

Robert Brown rcbrown at casbah.acns.nwu.edu
Mon Dec 16 14:57:13 EST 1991


Hi there,
	I am currently using in-situ hybridization to screen pcr products to
determine if any of the cDNAs are candidates for a receptor I am trying to
clone. Currently, I have gotten the technique to work in chicken brain, but
the tissue that I am really interested in is in the sheep pituitary (pars
tuberalis). The sense probes gave high background. I am interested in
conditions (washing, whether mercaptoethanol is used in washing solutions-I am
using s-35 probes, cpms/ml, specific activity of probes). I am also interested
to know if tissue can be stored at -70 degrees in OTC for any length of time.
I would appreciate any input I can get regarding this procedure.



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