On the website, it says
(Stock solution: 10 mg/ml in water (pH ≤6,0) or 0,01 M sodium acetate (pH 5,2). Do not heat RNase prepared in sodium acetate.)
(Working solution: 2-10 µg/ml in 10 mM Tris (pH 7,5), 15 mM NaCl.)
My real question is which temperature and for how much time should I heat prepare the stock solution( If I mix the powder in water)? while making the working solution from the stock solution in which quantity Ex- how much Tris and how much Nacl should I mix . This is my first time preparing Rnase A. So that is why I am a little bit confused
I think you can heat the stock solution (in water) at 100 °C for 5 to 15 min in order to eliminate all DNAse contaminations.
You probably already get it but I think the problem with sodium acetate is that acetate evaporates.
For your second question, it's just a recommanded dilution (10 mg/ml is too much) but it depends on your application that has the final word concerning the final concentration. Cited Tris and NaCl concentrations are also just recommandations. Some applications (eg. plasmid extraction) can propose some variations.
If you want to follow these recommandations, just prepare a sufficient amount of Tris-NaCl buffer (preferably with DNAse-free ingredients... such as autoclaved Tris 1M, NaCl 3M and milliq water) and make the dilution you want (2 or 10 µg/ml or whatever concentration you need) in the Tris-NaCl buffer with the recommanded concentrations.
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