Dear Flowers, We are trying to improve our assay for counting cells in 96 well plates using a fluorescent microplate reader. We are very lazy and skint (don't like buying kits) , and would like to add the dye to a well without the need to centrifuge and remove unbound dye. The characteristics of such a dye would be the following: 1. Fluorescence enhancement on binding to nucleic acid 2. High affinity for nucleic acid 3. DNA specificity. 4. Economic In the molecular probes catalogue we are told that Cyquant^TM green kit is the best dye for this purpose, and can detect down to 50 cells under optimal conditions, however we would like to find an alternative which is not a kit. We have tried PI + 1% Triton 100 which goes down to 2000 cells (no wash). Background fluorescence was very high for H33342 and DAPI (no wash). We were wondering whether we could should try one of the many new cyanine dyes. Which nucleic acid stains (other than Cyquant^TM green) would be best for the job? ?Thiazole orange ?TOTO / YOYO / BOBO/ HOHO^tm Many thanks for your patents. Robert Nordon Post Doctoral Fellow Graduate School of Biomedical Engineering University of New South Wales Sydney, 2052 Tel. no. 61-2-9385-1148 Fax 61-2-9663-2108
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