I am doing an experiment that requires me to fix some microbial cells, stain with antibodies, run through the flow cytometer and ultimately isolate DNA. The whole procedure is roughly 6 hours. Usually I fix cells for flow with PFA, but I'm concerned this may interfere with the DNA extraction and/or lower the DNA integrity. Can anyone suggest a more suitable fixative, something that wouldn't take more than 1hr?
Wolfgang Schechinger
Heating with excess tris can reverse aldehyde modifications of amino groups (imine formation), so I'd just try and set up a series of experiments e.g. perfoming PCRs or whatever you want to do with the DNA later.
Altenatives to PFA could be methanol or ethanol.
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