Pearls: 1. Be careful about stopping the pepsin reaction. Make sure that the pH is around 7 after you stop the reaction of pepsin by adding PBS. Sometimes, the pH after the first wash is still acidic and the pepsin keeps on working. 2. We have found that the PI solution stain should be increased 2X (from 50 ug/ml to 100 ug/ml). This improves the CV. Try it out and see if you agree. Hope you get some more Pearls. Dr. Timothy Kute, Ph.D. Associate Professor in Pathology Wake Forest University School of Medicine Winston-Salem, NC 27157-1073 Tel # 336-716-2950 Fax # 336-716-7595 E-mail tkute@wfubmc.edu -----Original Message----- From: Saldivar, Victor [mailto:victor_saldivar@srhc.iwhs.org] Sent: Thursday, June 15, 2000 6:11 PM To: cyto-inbox Subject: DNA ON PARAFFIN-EMBEDDED TISSUE--PLEASE POST Colleagues in FLOWCYTOMETRY: Every now and then we go back to having problems with obtaining good CVs (and reliable results) on deparaffinized tissue for DNA. My FLOW Supervisor requested to post th following question: "?Any (new) ideas about the steps to get the paraffin embedded tissue to a single cell suspension?". We will appreciate any "pearls". Victor A. Saldivar, MD -- San Antonio, TX <victor_saldivar@srhc.iwhs.org>
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