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RNAsin overdose! - (Feb/03/2010 )

Hi everyone,

I have a question about using RNAsin, at the DNase step.
In our protocol we should put 0.125 ul of RNAsin for each 1ul of DNAse, on each sample.
By mistake we put 10X more of RNAsin, but maintaing all the other products at their correct proportion in the mix.

We found out this mistake after putting 5ul (the recomended quantity) of this mix in each sample.
We would like to know the possibles future troubles this could bring to our material, and what could be the action of the presence of 10X more of RNAsin. Could it be inhibiting DNAse?
Thanks for your attention,
weŽll wait for your answers.

Laura, from my lab.


Hello everybody:

This problem is solved and if my experience could be useful, I just want to tell you the result. After making an agarose gel I could certify that there wasn’t any inhibition. Therefore, the reaction of DNAsa occurred correctly.

Thanks for all visitors.
See you soon :D .