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You will need to keep this in mind when analyzing the data. Assume that all of the -SH is modified by the mercurial. - The extinction coefficient for HSA is 0.53 per mg/ml per cm at 278 nm. - The quantum yield of HSA emission is 0.20 at pH 7.0, determined by comparison to tryptophan. - The refractive index of water is 1.33. - You may assume a value of 2/3 for the orientation factor. - General procedure: 1. Determine J Integrate HSAF from 311 to 450: 0.00991 cm Subtract HSAA from HSAMA. Convert difference to epsilon = A/(b*c) b=1 cm c=1e-5*0.58=5.8e-6 Mult epsilon*fluor*lambda^4*dlambda Sum up values; divide by area of fluorescence (0.00991) 2. Calculate Ro. 3. Calculate the energy transfer efficiency. 4. Calculate r, the inter-chromophore separation.