Dear Group, I was wondering if the above queston is actually true. The reason being that I had done some calculations regarding the final conc of PI in various protocols used for Viability and DNA analysis. and found that ususally for DNA 50ug/ml is the required conc ( for Flow- may be to maintain equilibrium in the stream etc but any ideas for fluorimetry???)and for viability it is around 1ug/ml. What is the reason? Do higher conc of PI in the staining buffer mean that more PI will enter the cells or is the cell totally impermeable to the PI. or that the PI conc used for viability has been standardized based on a concurrent experiment with say trypan blue or other methods? And how does time of incubation after adding the dye relate to the conc. After all both the methods are based on the same prinnciple that PI binds to DNA. Are there any simple proportionalities involved here eg more the dye less the time or mmore the dye more enters into the cells etc. Thanks in advance for all the tips. Rana Nagarkatti ____________________________________________________________________ Get free email and a permanent address at http://www.netaddress.com/?N=1
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