Figure 2.
Sequential steps of RBNS
(A) Purification of miRISC containing a single, unique small RNA guide.
(B) Total concentration of purified miRISC is estimated by northern blot.
(C) Concentration of binding-competent miRISC is measured by a titration experiment. KD is the apparent dissociation constant, r is the molar ratio of [RISC] to [RNA], n is the stoichiometric equivalence point, ƒ is the fraction bound, and ƒmax is the maximum fraction bound. Error bars report propagated SD.
(D) After the binding step of RBNS, the two membranes are separated, imaged, and quantified.