Date: Fri Jul 01 2005 - 14:18:23 EST
Dear Magda, You didn't provide information about the protocol you used, but I have several suggestions for you. 1. Use linear amplification, not log for DNA cell cycle analysis. 2. Check your protocol! The graph suggests that the cells were not properly permeabilized. 3. The first peak is not a G1 but are/were viable cells left unstained by the DNA dye. Was it PI? 4. The broad peak in the middle might be slightly damaged cells that took up some DNA dye. 5. The 2 peaks at the right of the graph probably represent G1 and G2 cells from a cell population in the sample that was already dead before you started. These cells are permeable to DNA stains. 6. Did you use cultured cells? 7. If so, I would like to recommend the Vindelov method (Cytometry 1983, March issue). Succes Willem Corver ________________________________ Van: Magda Lezcano [mailto:magda.lezcano@ebc.uu.se] Verzonden: wo 29-6-2005 21:38 Aan: Cytometry Mailing List Onderwerp: cell cycle protocol questions Dear followers, Im trying to do cell cycle analysis and I have got funny graphs, Im getting two picks at G2 and one of them is sometimes quite higher even than the G1, besides they are also the s-phase seems to be quite wide to me, so if anyone have any suggestion what could be my problem please let me know or if someone have any good protocol, please also forward it to me... thanks MAgda -- Magda Lezcano Department of Animal Development and Genetics EBC, Uppsala University Norbyvägen 18A, 75236 Uppsala, Sweden tel. +46-184712679 Fax. +46-184712683 mobile: 0735351834 "When joy is at its highest Sad thoughts run rife Youth and strength, how short they last How hopelessly we age!" Emperor Han Wudi, Western Han Dynasty
