rnase assay

[sunny]

can any one help me on bacterial rnase activity assay? which is the bst method for it?

[BabcockHall]

What is the specificity of the enzyme from bacteria, as opposed to ribonuclease A, for example? Which assays have you considered so far?

Edited February 12, 2013 by BabcockHall

[sunny]

helo

i used kamekura and onishi method for rnase activity.briefly its steps r:

buffer pH 7

enzyme .1ml

rna 1mg

incubation for 2 hr

stop solution

centrifuge

then in end dilution of supernatent.

in many other protocol more or less all steps are same but i cant get the logic related to dilutions.

is it the necessary step or what??? as if i have the O.D 2.06 of test y i dilute it????

and if i dilute at which dilution i used???

ooooh so confusing:^