Actinomycin D

[Ulrich Maier]

a03m at biologie.uni-bremen.de (Georg Kroeger) wrote:

>Dear Netters,

>we found an old box with actinomycin D in our fridge. We even got some
>bacteria, but how should we put them together. Could anyone be so kind
>and give us informations about how to dissolve it, and about useful
>concentrations for procaryotes!
>Thanks in advance!!

>Cheers

>Georg

You can dissolve Actinomycin D in pure Ethanol in a concentration of
5mg/ml as a stock solution. 

I have used it for human fibroblasts with a final concentration of
5µg/ml medium, but you´ll have to check the optimal concentration for
your assay by measuring the incorporation of 3H-uridine in the total
RNA after a short-time incubation of the bacteria with the
transcription inhibitor.
Actinomycin D blocks between 95 and 99% of overall transcription.    

Yours, U. Maier