Re-using hybridisation solutions containing double-stranded probes?

jfh jfhess at ucdavis.edu
Wed Mar 1 15:26:40 EST 1995


I agree, we reuse formamide probes.  Typical example is library
screening.  Save the initial hyb solution, Pick postives, plate out the
positives at the density you want for rescreening then reuse the hyb
solution to isolate pure plaques.

BTW, we do all hybs in formamide at 42 w/o any DNA in the hyb solution. 
Yes, that's right, southerns, northerns, lambdas, all in 5x sspe, 5 x D, 
0.5% SDS  50% form.

We've also been known to spike old hyb solution with fresh probe to bring
the cpms up. Just remember to heat the hyb solution before reuse.

John Hess, PhD                    Phone me 916 752 8420
Dept of Human Anatomy             FAX me 916 752 8520
University of Calif               Email me jfhess at ucdavis.edu
Davis, CA                         or leave me alone, your choice.



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