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Hey Daddy Jack,
With cunning recall, I remember the landmark Dunne paper describing a "sort-blot" technique where cells were sorted onto nitrocellulose for mRNA quantitation. We used this technique to confirm a more traditional approach of sorting lots of cells for a Northern analysis of heat shock protein mRNA in transformed CHO cells. Both approaches correlated quite well, with the Dunne sort-blot method requiring shorter sort times. The major problems I remember were handling the filter pieces, getting them to sit in the sort stream path and confining the sort-blot spot. Attempts to modify the procedure from autoradiography to a fluorescent probe we could measure on an image cytometer failed. If someone would like to try this promising, yet unrealized, technique I would be happy to rumage for old experimental notes.
-Bill
William Hyun, Director
Laboratory for Cell Analysis, Box 0808
University of California, San Francisco
San Francisco, CA 94143-0808
voice (415)476-2632
FAX (415)476-8218
email hyun@dmc.ucsf.edu
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web