Dear Charles, Probably the best method for DNA staining and sorting viable cell is the method that Margaret Goodell uses for the identification of SP cells. One place to get a very detailed procedure is Margaret's Website, http://www.bcm.tmc.edu/genetherapy/goodell/new_site/protocols.html . Modify the procedure in two simple ways; 1) always use verapamil to block Hoechst efflux, and 2) lower the PMT attenuation and gain so that G1 and G2M peaks are both on the histogram. Best Results, Jeffrey Jeffrey S. Haug Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, Missouri 64110 816-926-4142 FAX 816-926-2022 -----Original Message----- From: ckuszyns@UNMC.EDU [mailto:ckuszyns@UNMC.EDU] Sent: Monday, October 22, 2001 11:27 AM To: cyto-inbox Subject: Sorting on DNA Content Does anyone have a protocol for sorting cells based on DNA content. We have UV capabilities . Cells need to be viable following sorting.Thanks, Charles A. Kuszynski, Ph.D. Assistant Professor/Director Cell Analysis Facility University of Nebraska Medical Center 986495 Nebraska Medical Center Omaha, NE 68198-6495 402 559-6267 voice 402 559-4077 fax ckuszyns@unmc.edu
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